Lab 2: Gonadal function; FSH

LH clinical application - what does premature puberty do to gonadotropic secretion? what is the problem in a pituitary condition ? can lead to what? what is hypergonadotropic hypogonadism? what is hypogonadotropic hypogonadism and what happens to levels of GnRH, FSH, and LH?

-premature puberty: premature exposure to sex hormones; early gonadotropin secretion -lack of secretion by pituitary (pituitary conditions) -->dec LH can lead to infertility -->needs confirmation with other tests -hypergonadotropic hypogonadism: increase in LH -hypogonadotropic hypogonadism: decrease in LH and dec in hypothalamic conditions -->dec GnRH, dec FSH, dec LH -->needs confirmation with other tests

FSH clinical application - what happens to gonadotropin secretion in premature puberty? in females there is an increase in FSH during what 2 events?

-premature puberty: premature exposure to sex hormones; early gonadotropin secretion -in females, FSH increases during: -->Menopause -->Premature ovarian failure: >40mIU/mL; levels may be normalized with administration of estrogens (negative feedback mechanism)

what is the primary funciton of LH in males? what happens to LH levels during puberty? testosterone provides what for LH levels?

-primary function is to stimulate Leydig cells to produce testosterone -a normal increase in LH during puberty; testicular enlargement -testosterone provides negative feedback for LH levels

males FSH - regulates growth of ? maintenance of what process? what happens to FSH levels during puberty? acts on germinal stem cells and mature to ? do male hormones regulate (lower) FSH like females hormones do? what doe testosterone do with LH levels ?

-regulates growth of seminiferous tubules -maintenance of spermatogenesis -a normal increase in FSH during puberty; testicular enlargement -acts on germinal stem cells -->mature to spermatozoa through meiosis -male hormones do not regulate (lower) FSH like female hormones do; NO negative feedback -testosterone does have negative feedback but with LH

what sample do we use to test FSH? store at what temp for 48 hrs? if longer then freeze at what temp? can you freeze more than once? what are 3 interfering substances? what tube is used for collection?

-serum -->store samples at 2-8 C for up to 48 hrs -->freeze at -20 C for longer waiting periods (frozen only once); thawed an inverted before performing test -interfering substances: -->hemolyzed sample -->lipemic sample -->sodium additives: blood collection tubes (sodium azide) -EDTA (lavender) top tube

FSH - consists of what two subunits? what is special about the second subunit? how are levels of FSH produced/secreted controlled/regulated?

-alpha subunit: very similar in structure to other hormones: -->TSH, LH, HCG, and FSH -beta subunit: unique to FSH; biological and immunological properties -levels are controlled by sex hormones; negative feedback in females

LH clinical application - how to differentiate btw hypothalamic, pituitary or gonadal dysfunction? along with hormone levels, they are also used to determine what 3 things?

-differential diagnosis of hypothalamic, pituitary or gonadal dysfunction: -->FSH and LH assays -along with hormone levels, they are used to determine: -->menopause -->pinpoint ovulation -->monitor endocrine therapy

FSH clinical application - in males an increase in FSH can be caused by what 2 things with sperm? hypergonadotropic hypogonadism can be caused by failure of? can be from Klinefelter syndrome which has what symptoms? hypogonadotropin hypogonadism can be caused by what development in testicles? what is anorchia? low FSH can be caused by what 2 things also?

-increased FSH in males: -->azoospermia: absence of motile (viable) sperm -->oligospermia: low sperm count -Hypergonadotropic hypogonadism: increased FSH -->primary testicular failure: failure to produce sperm -->Klinefelter syndrome: common karyotype (XXY), small firm testicles, gynecomastia -Hypogonadotropin hypogonadism: decreased FSH -->abnormal testicle development -->anorchia (XY phenotype); born without testicles -->pituitary disease -->opioid use: dec GnRH (also dec in sperm motility and count, abnormal morphology)

what is FSH involved in controlling the growth and reproductive activities of? gonadal tissue synthesize?

-involved in the control of the growth of and reproductive activities of gonadal tissue -->testicles (males) -->ovaries (females) -gonadal tissue synthesizes and secretes male and female sex hormones

LH in females - what are the levels of progesterone and estradiol during luteal phase? this induces what process? if pregnancy occurs, the embryo produces what hormone? and the corpus luteum continues to produce what 2 hormones? if no pregnancy occurs what happens to the corpus luteum? what happens to levels of progesterone and estradiol

-luteal phase: high progesterone and second estradiol increase (negative feedback) -if pregnancy occurs: -->embryo produces HCG -->corpus luteum continues to produce progesterone and estradiol -if no pregnancy: -->corpus luteum degenerates -->decreased levels of progesterone and estradiol leads to menstruation

what are the expected values of FSH in males and females?

-males: 1.5-11.5 mIU/ml -females (for normal menstruating female): -->follicular phase: 3.2-10.0 mIU/ml -->mid cycle peak: 7.5-20.0 mIU/ml -->luteal phase: 1.3-11.0 mIU/ml -post menopause: 36.0-138.0 mIU/ml

diagnosis of hypogonadism - sample taken when is best for testosterone testing? in primary hypogonadism, there is a problem with? what is the level of FSH? in secondary hypogonadism there is a problem with? what is the FSH level? in tertiary hypogonadism there is a problem with? what is the FSH level?

-morning sample (8am-10am) for testosterone testing -FSH and LH testing: -->Primary hypogonadism: testes; FSH inc -->Secondary hypogonadism: pituitary; FSH normal or dec -->Tertiary hypogonadism: hypothalamic; FSH normal or dec

diagnosis of hypogonadism - sample taken when is best for testosterone testing? in primary hypogonadism, there is a problem with? what is the level of LH? in secondary hypogonadism there is a problem with? what is the LH level? in tertiary hypogonadism there is a problem with? what is the LH level?

-morning sample (8am-10am) for testosterone testing -FSH and LH testing: -->Primary hypogonadism: testes; LH inc -->Secondary hypogonadism: pituitary; LH normal or dec -->Tertiary hypogonadism: hypothalamic; LH normal or dec

lab assay; ELISA (sandwich ELISA) - in a sandwich ELISA the wells are coated with what? what is the HRP conjugate? FSH is sandwiched btw what two antibodies? unbound labeled anti-B-FSH antibody are? what is the substrate and what color is it? what color does stop solution turn the reaction? absorbance is at? the intensity of the color is what to FSH concentration?

-wells coated with mouse monoclonal anti-alpha FSH (solid phase) -FSH is sandwiched btw solid phase and HRP labeled antibodies -unbound labeled anti-Beta FSH antibodies are washed -TMB (substrate) added (blue color) -stop solution (1N HCL), stops color development (yellow color) -absorbance at 450 nm -intensity of color is directly proportional to FSH concentration

LH in females - what happens to estradiol and progesterone levels when menstrual cycle begins (day 1)? as a result what does the hypothalamus do with GnRH and what does the anterior pituitary do with FSH? as FSH increases what happens to follicles? what is being secreted also? on days 12-14 what happens to LH when estradiol rise occurs? when does ovulation occur? egg is released and what is formed? this secretes what 2 sex hormones?

-Day 1 (menstrual cycle begins: decreased estradiol and progesterone -->hypothalamus releases GnRH which stimulates anterior pituitary -->anterior pituitary releases FSH -->as FSH increases, several follicles are stimulated (follicular phase) -->as follicle develops, estradiol is being secreted -Day 12-13 (fast increase with peak at day 14): -->LH is released when this estradiol rise happens -->ovulation happens 12-18 hrs after LH peaks -->egg is released and corpus luteum is formed; secretes progesterone and estrogen (negative feedback of LH)

in males, do testicular tumors increase or decrease FSH? do testicular tumors increase or decrease LH?

-FSH: decrease -LH: increase

what is FSH ? what is it secreted by ? what is GnRH? where is it secreted from? control release of what two hormones?

-Follicle stimulating hormone (FSH) is a glycoprotein -->secreted by basophilic cells of the anterior pituitary -Gonadotropin releasing hormone (GnRH) -->is secreted from the hypothalamus -->control release of FSH and LH

FSH clinical application - what is hypergonadotropic hypogonadism (what happens to FSH)? when does this occur in females that is not due to disease? what is hypogonadotropin hypogonadism (what happens to FSH)? seen with secondary?

-Hypergonadotropic hypogonadism: increased FSH -->ovarian failure/menopause (normal; premature before 40 years old -Hypogonadotropin hypogonadism: decreased FSH -->secondary amenhorrhea: anorexia, intense physical exercise, prolactin production, pituitary tumors

LH clinical application - in females, inc LH occurs when? what is hypergonadotropic hypogonadism (what happens to LH)? when does this occur in females that is not due to disease? what is hypogonadotropin hypogonadism (what happens to LH)? seen with secondary?

-Hypergonadotropic hypogonadism: increased LH -->ovarian failure/menopause (normal; premature before 40 years old -Hypogonadotropin hypogonadism: decreased LH -->secondary amenorrhea: anorexia, intense physical exercise, prolactin production, pituitary tumors

LH clinical application - hypergonadotropic hypogonadism can be due to failure of? can be due to Klinefelter syndrome which has what symptoms? hypogonadotropin hypogonadism can be caused by what development in testicles? what is anorchia? can be caused by what 2 things?

-Hypergonadotropic hypogonadism: increased LH -->primary testicular failure: failure to produce sperm -->Klinefelter syndrome: common karyotype (XXY), small firm testicles, gynecomastia -Hypogonadotropin hypogonadism: decreased LH -->abnormal testicle development -->anorchia (XY phenotype); born without testicles -->pituitary disease -->opioid use: dec GnRH (also dec in sperm motility and count, abnormal morphology)

what is LH? produced by what ? in response to? consists of what subunits? levels are controlled by ?

-Luteinizing hormone (LH) is a glycoprotein -produced by anterior pituitary in response to GnRH released by the hypothalamus -alpha subunit: very similar in structure to: -->TSH, LH, HCG, and FSH -beta subunit: unique; biological and immunological properties -levels are controlled by sex hormones (negative feedback)

what are the expected values of LH in males and females?

-Males: 1.2-7.8mIU/mL -Females: -->Follicular phase: 1.7-15.0mIU/mL -->Ovulatory peak: 21.9-56.6mIU/mL -->Luteal phase: 0.6-16.3mIU/mL -Post-menopausal: 14.2-52.3mIU/mL

females FSH - what happens to FSH levels during puberty? stimulates growth and maturation of ? promotes follicular steroidogenesis of what two hormones? stimulates LH production which does what? increased estradiol production occurs as? stimulates what to happen to FSH receptor activity? this is needed for maturation of what?

-a normal increase in FSH during puberty -stimulates growth and maturation of ovarian follicles -promotes follicular steroidogenesis -->progesterone -->estrogen -simulates LH (luteinizing hormone) production -->which binds to theca cells of ovaries to simulate steroidogenesis -increased estradiol production occurs as follicular maturation progresses -->stimulates increased FSH receptor activity and follicular binding; which is needed for egg maturation

LH in females - what happens to LH during puberty? concentration depends on what?

-a normal increase in LH during puberty -variation in concentration depends on the menstrual cycle

reagents - all reagents should be at what temp before use? what should be done with reagents before use? reagents come how? what is coating the microtiter wells? what is the HRP labeled conjugate reagent? how many standards are used? what are 3 limitations of the procedure? what does inadequate washing lead to false levels of?

-all reagents should be brought to room temp before use -all regents should be mixed by gentle inversion or swirling prior to use; do not induce foaming -reagents come lyophilized and need to be reconstituted -mouse monoclonal anti-alpha FSH coated microtiter wells -HRP labeled conjugate reagent: mouse monoclonal anti-Beta FSH -6 FSH standards (mIU/mL) -limitations: -->wash procedure is critical; inadequate washing can lead to false elevated readings in abs -->lipemic, hemolyzed, turbid samples -->additives