FSH Porcine

Porcine Follicle Stimulating Hormone
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Description

Definition and Source of FSH Porcine

Follicle-Stimulating Hormone Porcine (FSH Porcine) is a glycoprotein hormone derived from porcine pituitary glands. It consists of α and β subunits, with the β subunit conferring biological specificity . The hormone is purified through proprietary chromatographic techniques to achieve high biological activity (40 IU/mg) , making it critical for reproductive research and veterinary applications.

Biological Functions

FSH Porcine regulates reproductive processes in mammals through two primary mechanisms:

  • In females: Stimulates follicular growth by targeting granulosa cells, promotes progesterone synthesis via autophagy-mediated lipid droplet degradation , and modulates aquaporin expression in ovarian tissues .

  • In males: Enhances Sertoli cell proliferation and androgen-binding protein production, supporting spermatogenesis .

Superovulation in Cattle

FSH Porcine is commercially formulated as FOLLTROPIN®, which induces superovulation in dairy and beef cows. Key protocols include:

ParameterDetail
Dosage87.5 IU twice daily for 4 days (intramuscular)
Prostaglandin co-adminCloprostenol or dinoprost tromethamide with the 6th FSH dose
Embryo collection7 days post-insemination

In Vitro Studies

  • Ovarian follicle culture: Porcine FSH (10–100 mIU/mL) improves antral follicle development and oocyte maturation in goats .

  • Cell signaling: Activates PI3K/AKT and SAPK/JNK pathways in granulosa cells, upregulating BECLIN1 expression to enhance autophagy and progesterone synthesis .

Molecular Mechanisms in Granulosa Cells

  • Autophagy induction: FSH Porcine increases LC3-II levels and autophagic vacuole formation, accelerating lipid droplet breakdown to fuel progesterone production .

  • Pathway regulation:

    • PI3K/AKT inhibition reduces FSH-mediated BECLIN1 upregulation by 65% .

    • JNK/c-Jun pathway blockade decreases progesterone synthesis by 40% .

Sertoli Cell Proliferation

  • Boar FSH content peaks at 60 days postnatal, correlating with Sertoli cell proliferation .

  • Optimal in vitro proliferation occurs at 75 ng/mL FSH (72-hour exposure) .

Comparative Efficacy of FSH Sources

ParameterPorcine FSHRecombinant Bovine FSHRecombinant Human FSH
Bioactivity (IU/mg)150–170 7,000 9.4–14.2
Oocyte maturation rate58% (antral) 62% 55%
Steroidogenic gene modulationCYP17, ↑ CYP19A1 Minimal effect Moderate effect

Physicochemical Properties and Handling

PropertySpecification
Solubility≥100 µg/mL in sterile H₂O
Storage (lyophilized)-18°C (stable for 3 weeks at 25°C)
Reconstituted stability2–7 days at 4°C; long-term storage at -80°C

Industrial and Recombinant Production

  • Yeast expression: Codon optimization and HSA fusion strategies increase recombinant pFSH yields to 40.8 mg/L in Pichia pastoris .

  • Biological activity: Recombinant pFSH stimulates cAMP synthesis in HEK293 cells expressing porcine FSHR .

Product Specs

Introduction
Follicle-stimulating hormone (FSH) is crucial for reproductive function in both males and females. It's produced by the anterior pituitary gland. In females, FSH promotes the growth and maturation of ovarian follicles, essential for egg development. In males, FSH is indispensable for sperm production, supporting the function of Sertoli cells in the testes. Both genders rely on FSH for germ cell maturation.
Description
This product consists of purified porcine FSH, derived from pig pituitary glands and meticulously processed through advanced chromatographic techniques.
Physical Appearance
The product appears as a sterile, white powder resulting from a freeze-drying process.
Formulation
The FSH product is freeze-dried without any additional ingredients.
Solubility
For optimal reconstitution, dissolve the freeze-dried porcine FSH in sterile 18M-cm H2O at a concentration not lower than 100 µg/ml. Further dilutions can be made using other aqueous solutions.
Stability
While the freeze-dried porcine FSH remains stable for 3 weeks at room temperature, it's best stored in a dry environment below -18°C. After reconstitution, keep it at 4°C for a maximum of 2-7 days. For long-term storage, add a carrier protein (0.1% HSA or BSA) and store below -18°C. Avoid repeated freezing and thawing.
Specific Activity

The product boasts a specific activity of 40 Units per 1 mg.

Synonyms
Follitropin, Follicle-stimulating hormone, FSH.
Source
Porcine pituitaries.

Q&A

What is the physiological role of FSH in porcine reproduction?

FSH plays a critical role in controlling follicular growth and maturation in the porcine ovary under physiological conditions. It binds to its receptor exclusively on the surface of granulosa cells to stimulate the PI3K/AKT signaling pathway . In commercial applications, exogenous hormones including FSH are used to control follicular development and synchronize ovulation in the batch flow management of gilts and weaned sows, which improves economic benefits in swine production .

Unlike simple reproductive stimulation, FSH initiates complex cascades of molecular events. The hormone activates multiple pathways including PI3K/AKT and SAPK/JNK, which subsequently regulate gene expression related to follicular development, maturation, and ovulation . These gene expression changes drive physiological outcomes such as ovarian weight gain, follicular development, and ultimately successful ovulation and fertility.

How does porcine FSH differ from other mammalian FSH variants?

Porcine FSH exhibits species-specific characteristics while maintaining the core conserved structure of glycoprotein hormones. Research focusing on species variations is important because heterologous proteins like equine chorionic gonadotropin (eCG) can generate antibodies when used across species, as observed in cattle superovulation protocols .

When designing experiments with porcine FSH, researchers should consider species-specific binding affinities and downstream signaling cascades. Protocols developed for bovine or human FSH may require significant adjustments when applied to porcine models. This is particularly relevant when developing recombinant versions of porcine FSH, where protein structure and post-translational modifications must closely mimic the native hormone to maintain bioactivity and reduce immunogenicity.

What experimental models are most appropriate for porcine FSH research?

The selection of appropriate experimental models depends on the specific research questions. For pharmacokinetic studies, both rat and sow models have been successfully used to evaluate the half-life of recombinant porcine FSH variants . For in vitro analyses of FSH activity, isolated porcine follicles containing granulosa cells provide valuable insights into cellular and molecular mechanisms.

When isolating porcine follicles, researchers should follow established protocols: obtain ovaries from local abattoirs, transport them in sterile physiological saline at 33-35°C, wash them with saline containing antibiotics, and process the ovarian cortex into appropriate sections . Only follicles with clear, centrally located oocytes and healthy granulosa cells should be selected for experiments, with typical follicular diameters ranging from 150-250μm at the beginning of culture .

How can researchers develop and evaluate recombinant porcine FSH variants?

The development of recombinant porcine FSH involves protein fusion technology to engineer molecules with enhanced pharmacological properties. A successful approach has been to create a long-acting recombinant porcine FSH (rpFSH-pFc) by combining porcine FSH with porcine fragment crystallizable (Fc) via a (G4S)3 linker . This fusion strategy increases the half-life of the hormone while maintaining its biological activity.

Evaluation of recombinant FSH variants requires multiple experimental approaches:

  • Pharmacokinetic studies in both small animal models (rats) and target species (sows) to determine half-life

  • In vitro functional assays including cAMP level evaluation and germinal vesicle breakdown (GVBD) analysis

  • In vivo efficacy testing through ovarian weight gain, superovulation, and fertility testing assays

  • Molecular analysis of target gene expression related to follicular development, maturation, and ovulation

These comprehensive evaluations ensure that the recombinant protein maintains biological activity comparable to native FSH while offering improved pharmacological properties such as extended half-life.

What signaling pathways mediate FSH action in porcine granulosa cells?

FSH activates multiple signaling pathways in porcine granulosa cells, with the PI3K/AKT and SAPK/JNK pathways being particularly important. Contrary to initial hypotheses that FSH might inhibit autophagy through AKT-mediated activation of mTOR, research has demonstrated that FSH actually induces autophagy in porcine granulosa cells .

The molecular mechanism involves FSH binding to its receptor, activating PI3K/AKT, which then phosphorylates the transcription factor c-Jun through the SAPK/JNK pathway. Phosphorylated c-Jun translocates into the nucleus and binds to the BECLIN1 promoter region, increasing Beclin1 expression and enhancing autophagy . This pathway can be experimentally verified using specific inhibitors:

  • LY294002 (PI3K/AKT inhibitor) blocks FSH-induced increases in Beclin1 levels

  • SP600125 (SAPK/JNK inhibitor) prevents the increase in Beclin1 levels induced by FSH

  • Knockdown of c-Jun prevents FSH-induced increases in Beclin1 expression

Understanding these signaling cascades is critical for designing targeted interventions and for interpreting experimental results in reproductive biology research.

How does FSH regulate autophagy in porcine granulosa cells?

FSH upregulates autophagy in porcine granulosa cells through a specific molecular pathway that can be experimentally demonstrated through multiple approaches. The autophagy marker LC3-II increases in a dose-dependent manner with FSH treatment (optimal at 0.01 IU/mL), and electron microscopy reveals increased autophagic vacuoles in granulosa cells after FSH stimulation .

To properly assess FSH-induced autophagy, researchers should employ:

  • Western blotting to detect LC3-II levels at various FSH concentrations and time points

  • LC3 turnover assay using chloroquine to block autolysosome degradation

  • Electron microscopy to visualize and quantify autophagic vacuoles

  • Gene expression analysis of key autophagy regulators like Beclin1

It's important to note that contrasting observations have been reported in mouse granulosa cells, where FSH appeared to inhibit autophagy through AKT-mediated activation of mTOR . These species-specific differences highlight the importance of using appropriate animal models and verifying mechanisms through multiple experimental approaches.

What are the optimal conditions for in vitro culture of porcine follicles and granulosa cells?

For successful in vitro culture of porcine follicles and granulosa cells, researchers should adhere to specific protocols:

  • Collection and Processing:

    • Obtain ovaries from local abattoirs and transport to the laboratory in sterile physiological saline (33-35°C) within 2-3 hours

    • Wash ovaries twice with sterile physiological saline containing antibiotics (100 IU/mL penicillin and 50 mg/mL streptomycin)

    • Cut ovarian cortex into sections approximately 500-μm thick and cross-chop into 1mm × 1mm pieces

  • Follicle Selection:

    • Select only follicles containing a clear, centrally located oocyte and healthy granulosa cells

    • Typical follicular diameter should be 150-250μm at the beginning of culture

  • Culture Conditions:

    • Culture individually in 96-well plates containing DMEM/F12 supplemented with 3 mg/mL BSA

    • Maintain in humidified air with 5% CO₂

    • For FSH experiments, use concentrations between 0.001-0.1 IU/mL, with 0.01 IU/mL being optimal for most applications

Following these protocols ensures reproducible results and maintains cellular viability and function during experimental manipulations.

How should researchers design pharmacokinetic studies for recombinant porcine FSH?

Pharmacokinetic studies of recombinant porcine FSH require careful experimental design to accurately determine half-life and bioavailability. A comprehensive approach should include:

  • Multiple animal models:

    • Small animal models (rats) for initial screening and dose-finding

    • Target species (sows) for confirming translational relevance

  • Sampling protocols:

    • Collect blood samples at regular intervals post-administration

    • Process samples consistently to minimize variation

    • Use validated assays to measure FSH levels in serum

  • Comparative analysis:

    • Include native porcine FSH as a reference standard

    • Test multiple formulations simultaneously when possible

    • Analyze data using appropriate pharmacokinetic models

These studies should determine key parameters such as absorption rate, distribution volume, elimination half-life, and area under the curve (AUC). For long-acting recombinant versions like rpFSH-pFc, extended sampling periods are necessary to capture the prolonged activity profile characteristic of Fc-fusion proteins .

What techniques are most effective for analyzing molecular responses to FSH?

Multiple molecular analysis techniques should be employed to comprehensively evaluate FSH responses:

  • Protein expression analysis:

    • Western blotting for detecting key proteins like LC3-II, Beclin1, and phosphorylated signaling molecules (p-AKT, p-JNK, p-c-Jun)

    • Use appropriate loading controls (β-actin, Histone 3) and normalize expression levels

  • Gene expression analysis:

    • RT-PCR or qPCR for measuring mRNA levels of target genes associated with follicular development, maturation, and ovulation

    • Include reference genes for normalization

  • Functional assays:

    • cAMP measurement to assess receptor activation

    • Germinal vesicle breakdown (GVBD) analysis for oocyte maturation

    • Progesterone secretion assays to evaluate steroidogenic responses

  • Visualization techniques:

    • Electron microscopy for autophagic vacuoles

    • Fluorescent staining (BODIPY 493/503) for lipid droplets

    • Immunofluorescence for protein localization

Combining these approaches provides a comprehensive understanding of the molecular mechanisms underlying FSH action in porcine reproductive tissues.

How can researchers quantitatively assess FSH efficacy in porcine models?

Quantitative assessment of FSH efficacy requires multiple endpoints that span molecular, cellular, and physiological levels:

  • Molecular markers:

    • Upregulation of target genes associated with follicular development

    • Activation of key signaling pathways (PI3K/AKT, SAPK/JNK)

    • Expression of steroidogenic enzymes

  • Cellular responses:

    • cAMP production in granulosa cells

    • Autophagy induction measured by LC3-II levels

    • Lipid droplet metabolism in steroidogenic cells

  • Physiological outcomes:

    • Ovarian weight gain

    • Number and quality of developed follicles

    • Superovulation response

    • Fertility rates following artificial insemination

Data should be analyzed using appropriate statistical methods, with results typically presented as mean ± standard deviation or standard error. Significant differences between experimental groups can be determined using t-tests for pairwise comparisons or ANOVA for multiple group comparisons, with p-values less than 0.05 considered statistically significant .

How should researchers address contradictory findings in FSH research?

When confronted with contradictory findings, researchers should follow these approaches:

  • Consider species differences:

    • FSH appears to induce autophagy in porcine granulosa cells but may inhibit it in mouse granulosa cells

    • These differences may reflect genuine biological variations rather than experimental errors

  • Examine methodological differences:

    • Variation in experimental conditions (FSH concentration, timing, culture conditions)

    • Differences in readout methods or endpoints

    • Use of inhibitors or genetic manipulations that may have off-target effects

  • Validate with multiple techniques:

    • Confirm autophagy induction using both LC3-II western blotting and electron microscopy

    • Validate signaling pathway involvement using both pharmacological inhibitors and genetic approaches

    • Measure functional outcomes alongside molecular markers

For example, when investigating whether FSH induces or inhibits autophagy, researchers should conduct comprehensive autophagic flux assays rather than relying solely on static LC3-II levels, as was done to confirm that FSH enhances autophagy in porcine granulosa cells .

What statistical approaches are most appropriate for FSH experimental data?

Statistical analysis of FSH experimental data should be tailored to the specific experimental design and data characteristics:

When reporting statistical significance, use consistent notation (e.g., *, P < 0.05; **, P < 0.01) and specify the statistical tests employed. Sample sizes should be determined through power analysis when possible, and biological replicates should be distinguished from technical replicates .

What are the prospects for developing improved recombinant porcine FSH variants?

The development of long-acting recombinant porcine FSH (rpFSH-pFc) represents a significant advancement, but several directions for further improvement exist:

  • Optimization of fusion partners:

    • Exploring alternative protein fusion technologies beyond Fc fusion

    • Fine-tuning linker sequences to balance stability and bioactivity

    • Investigating site-specific modifications to enhance receptor binding

  • Glycosylation engineering:

    • Controlling glycosylation patterns to optimize half-life and bioactivity

    • Exploring glycoengineered expression systems to produce more consistent glycoforms

    • Evaluating the immunogenicity of different glycosylation profiles

  • Formulation development:

    • Creating sustained-release formulations for even longer duration of action

    • Developing temperature-stable preparations for field applications

    • Exploring alternative delivery routes beyond injection

These improvements could address current limitations while maintaining the advantages of species-specific recombinant proteins over heterologous hormones like eCG, which can cause side effects including follicular cysts, premature luteinization, and antibody generation .

How does FSH-induced autophagy interact with other cellular processes in porcine granulosa cells?

The discovery that FSH promotes progesterone production by enhancing autophagy opens several new research questions:

  • Interaction with lipid metabolism:

    • FSH reduces the number and size of lipid droplets in porcine granulosa cells

    • This effect is dependent on autophagy genes (ATG5, BECLIN1)

    • Further research is needed to elucidate how lipid droplet degradation through autophagy contributes to steroidogenesis

  • Cross-talk with classical steroidogenic pathways:

    • FSH promotes progesterone production through both autophagy-dependent mechanisms and by increasing steroidogenic enzyme expression

    • The relative contribution of each pathway under different physiological conditions requires investigation

    • Understanding how these pathways integrate could lead to more effective reproductive interventions

  • Role in follicular development beyond steroidogenesis:

    • Autophagy may influence granulosa cell proliferation, differentiation, and survival

    • The impact of FSH-induced autophagy on follicular selection and dominance needs exploration

    • Long-term consequences for oocyte quality and developmental competence remain to be determined

These research directions could provide a more comprehensive understanding of FSH action in the ovary and identify new therapeutic targets for reproductive disorders.

What methodological adaptations are needed when translating porcine FSH research to other species?

Translating findings from porcine FSH research to other species requires careful methodological considerations:

  • Species-specific receptor binding:

    • Evaluate receptor binding affinities and activation profiles across species

    • Consider developing species-specific recombinant FSH variants

    • Adjust dosing based on receptor sensitivity rather than simple body weight conversion

  • Signaling pathway conservation:

    • Confirm whether the PI3K/JNK/c-Jun pathway mediating FSH-induced autophagy is conserved across species

    • Adapt inhibitor concentrations based on species-specific sensitivity

    • Validate key molecular interactions (e.g., c-Jun binding to BECLIN1 promoter) in target species

  • Physiological endpoint relevance:

    • Consider species differences in follicular development patterns

    • Adjust timing of measurements based on species-specific reproductive cycles

    • Develop appropriate biomarkers for follicular development that are relevant across species

These adaptations ensure that research findings maintain translational relevance while acknowledging the biological diversity across mammalian reproductive systems.

Product Science Overview

Structure and Function

Follicle Stimulating Hormone (FSH) is composed of two subunits: alpha (α) and beta (β). The alpha subunit is common to other glycoprotein hormones like luteinizing hormone (LH), thyroid-stimulating hormone (TSH), and human chorionic gonadotropin (hCG). The beta subunit, however, is unique to FSH and confers its specific biological activity .

In female pigs, pFSH stimulates the growth and maturation of ovarian follicles, which are essential for ovulation and fertility. It works in concert with luteinizing hormone (LH) to regulate the menstrual cycle and reproductive processes. In male pigs, pFSH is involved in the regulation of spermatogenesis, the process by which sperm is produced .

Biological Significance

The biological significance of pFSH extends beyond its role in reproduction. It has been found that overexpression of pFSH can lead to an improvement in female fecundity. Studies have shown that pituitary-specific overexpression of pFSH in transgenic mice leads to an increase in ovulation rate and litter size without causing reproductive defects . This indicates that pFSH plays a significant role in enhancing reproductive efficiency and could be a valuable tool in animal breeding programs.

Applications in Research and Medicine

Porcine FSH is widely used in scientific research to study reproductive physiology and endocrinology. It is also used in veterinary medicine to treat reproductive disorders in pigs. The recombinant form of pFSH is used in various experimental setups to understand its role in follicular development and hormone regulation .

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