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Multiple luteinizing hormone receptor (LHR) protein variants, interspecies reactivity of anti-LHR mAb clone 3B5, subcellular localization of LHR in human placenta, pelvic floor and brain, and possible role for LHR in the development of abnormal pregnancy, pelvic floor disorders and Alzheimer's disease.

Bukovsky A, Indrapichate K, Fujiwara H, Cekanova M, Ayala ME, Dominguez R, Caudle MR, Wimalsena J, Elder RF, Copas P, Foster JS, Fernando RI, Henley DC, Upadhyaya NB - Reprod. Biol. Endocrinol. (2003)

Bottom Line: Besides ovaries, LHR immunoreactivity (LHRI) was also found in other tissues, such as the brain, fallopian tube, endometrium, trophoblast and resident tissue macrophages.In normal human term placentae, villous LHRI was associated with blood sinusoids and cytotrophoblast cells, and rarely detected in trophoblastic syncytium.Our observations indicate that, in contrast to the theory on the role of vascular hormone receptors in preferential pick up of circulating hormones, there is no need to enhance selective pick up rather only prevent LH/CG transport to inappropriate sites.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratory for Development, Differentiation and Cancer, Department of Obstetrics and Gynecology, The University of Tennessee Graduate School of Medicine, 1924 Alcoa Highway, Knoxville, TN 37920, USA. buko@utk.edu

ABSTRACT
Distinct luteinizing hormone receptor (LHR) protein variants exist due to the posttranslational modifications. Besides ovaries, LHR immunoreactivity (LHRI) was also found in other tissues, such as the brain, fallopian tube, endometrium, trophoblast and resident tissue macrophages. The 3B5 mouse monoclonal antibody was raised against purified rat LHR. In rat, porcine and human ovaries, the 3B5 identified six distinct LHR bands migrating at approximately 92, 80, 68, 59, 52 and 48 kDa. Characteristic LHRI was detected in rat, human and porcine corpora lutea. During cellular differentiation, subcellular LHR distribution changed from none to granular cytoplasmic, perinuclear, surface, nuclear and no staining. There were also differences in vascular LHR expression--lack of LHRI in ovarian vessels and strong staining of vessels in other tissues investigated. In normal human term placentae, villous LHRI was associated with blood sinusoids and cytotrophoblast cells, and rarely detected in trophoblastic syncytium. In all abnormal placentae, the LHRI of sinusoids was absent, and syncytium showed either enhanced (immature placental phenotypes) or no LHRI (aged placental phenotype). LHRI in human brain was identified in microglial cells (CD68+ resident macrophages). Protein extracts from human vaginal wall and levator ani muscle and fascia showed strong approximately 92 and 68 kDa species, and LHRI was detected in smooth muscle cells, fibroblasts, resident macrophages and nuclei of skeletal muscle fibers. Our observations indicate that, in contrast to the theory on the role of vascular hormone receptors in preferential pick up of circulating hormones, there is no need to enhance selective pick up rather only prevent LH/CG transport to inappropriate sites. Abnormal placental LHR expression may play a role in the development of abnormal pregnancy. Expression of LHR in the pelvic floor compartments suggests that high LH levels in postmenopausal women may contribute to the pelvic floor relaxation and increased incidence of pelvic floor disorders. Since chorionic gonadotropin increases secretion of a variety of cytokines by monocytes, and induces their inflammatory reaction and phagocytic activity, high LH levels in aging individuals may also activate microglia (mononuclear phagocyte system in the central nervous system) and contribute to the development of Alzheimer's disease and other inflammation-mediated neurodegenerative diseases.

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LHR expression in the rat ovaries and uterus. A) Strong LHR immunoreactivity is apparent in mature (MCL) and regressing CL (RCL) and interstitial cells (i). Note unstained vessels (v). B) Strong staining of theca interna (t) and weaker staining of mature granulosa cells (mg) in preovulatory follicle. Note scarce cytoplasmic granules with LHR immunoreactivity in immature granulosa cells (ig). C) Control. D) Increased staining during granulosa cell differentiation (dg) in large antral follicle. E) In the rat uterus, the LHR immunostaining is apparent in the surface epithelium (se), uterine glands (ug) and vasculature (v). Uterine stromal cells show nuclear staining (arrowhead). No hematoxylin counterstain.
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Figure 1: LHR expression in the rat ovaries and uterus. A) Strong LHR immunoreactivity is apparent in mature (MCL) and regressing CL (RCL) and interstitial cells (i). Note unstained vessels (v). B) Strong staining of theca interna (t) and weaker staining of mature granulosa cells (mg) in preovulatory follicle. Note scarce cytoplasmic granules with LHR immunoreactivity in immature granulosa cells (ig). C) Control. D) Increased staining during granulosa cell differentiation (dg) in large antral follicle. E) In the rat uterus, the LHR immunostaining is apparent in the surface epithelium (se), uterine glands (ug) and vasculature (v). Uterine stromal cells show nuclear staining (arrowhead). No hematoxylin counterstain.

Mentions: We also included absorption studies with porcine ovarian homogenates. Primary antibody (100 μg) in 2 ml PBS was incubated 20 min with 500 mg of porcine ovarian homogenate (late follicular phase) at room temperature. After centrifugation (11000 × g for 10 min at 4°C), the supernatant was diluted to 10 ml with blocking reagent and used for immunostaining of nitrocellulose membranes.


Multiple luteinizing hormone receptor (LHR) protein variants, interspecies reactivity of anti-LHR mAb clone 3B5, subcellular localization of LHR in human placenta, pelvic floor and brain, and possible role for LHR in the development of abnormal pregnancy, pelvic floor disorders and Alzheimer's disease.

Bukovsky A, Indrapichate K, Fujiwara H, Cekanova M, Ayala ME, Dominguez R, Caudle MR, Wimalsena J, Elder RF, Copas P, Foster JS, Fernando RI, Henley DC, Upadhyaya NB - Reprod. Biol. Endocrinol. (2003)

LHR expression in the rat ovaries and uterus. A) Strong LHR immunoreactivity is apparent in mature (MCL) and regressing CL (RCL) and interstitial cells (i). Note unstained vessels (v). B) Strong staining of theca interna (t) and weaker staining of mature granulosa cells (mg) in preovulatory follicle. Note scarce cytoplasmic granules with LHR immunoreactivity in immature granulosa cells (ig). C) Control. D) Increased staining during granulosa cell differentiation (dg) in large antral follicle. E) In the rat uterus, the LHR immunostaining is apparent in the surface epithelium (se), uterine glands (ug) and vasculature (v). Uterine stromal cells show nuclear staining (arrowhead). No hematoxylin counterstain.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC161821&req=5

Figure 1: LHR expression in the rat ovaries and uterus. A) Strong LHR immunoreactivity is apparent in mature (MCL) and regressing CL (RCL) and interstitial cells (i). Note unstained vessels (v). B) Strong staining of theca interna (t) and weaker staining of mature granulosa cells (mg) in preovulatory follicle. Note scarce cytoplasmic granules with LHR immunoreactivity in immature granulosa cells (ig). C) Control. D) Increased staining during granulosa cell differentiation (dg) in large antral follicle. E) In the rat uterus, the LHR immunostaining is apparent in the surface epithelium (se), uterine glands (ug) and vasculature (v). Uterine stromal cells show nuclear staining (arrowhead). No hematoxylin counterstain.
Mentions: We also included absorption studies with porcine ovarian homogenates. Primary antibody (100 μg) in 2 ml PBS was incubated 20 min with 500 mg of porcine ovarian homogenate (late follicular phase) at room temperature. After centrifugation (11000 × g for 10 min at 4°C), the supernatant was diluted to 10 ml with blocking reagent and used for immunostaining of nitrocellulose membranes.

Bottom Line: Besides ovaries, LHR immunoreactivity (LHRI) was also found in other tissues, such as the brain, fallopian tube, endometrium, trophoblast and resident tissue macrophages.In normal human term placentae, villous LHRI was associated with blood sinusoids and cytotrophoblast cells, and rarely detected in trophoblastic syncytium.Our observations indicate that, in contrast to the theory on the role of vascular hormone receptors in preferential pick up of circulating hormones, there is no need to enhance selective pick up rather only prevent LH/CG transport to inappropriate sites.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratory for Development, Differentiation and Cancer, Department of Obstetrics and Gynecology, The University of Tennessee Graduate School of Medicine, 1924 Alcoa Highway, Knoxville, TN 37920, USA. buko@utk.edu

ABSTRACT
Distinct luteinizing hormone receptor (LHR) protein variants exist due to the posttranslational modifications. Besides ovaries, LHR immunoreactivity (LHRI) was also found in other tissues, such as the brain, fallopian tube, endometrium, trophoblast and resident tissue macrophages. The 3B5 mouse monoclonal antibody was raised against purified rat LHR. In rat, porcine and human ovaries, the 3B5 identified six distinct LHR bands migrating at approximately 92, 80, 68, 59, 52 and 48 kDa. Characteristic LHRI was detected in rat, human and porcine corpora lutea. During cellular differentiation, subcellular LHR distribution changed from none to granular cytoplasmic, perinuclear, surface, nuclear and no staining. There were also differences in vascular LHR expression--lack of LHRI in ovarian vessels and strong staining of vessels in other tissues investigated. In normal human term placentae, villous LHRI was associated with blood sinusoids and cytotrophoblast cells, and rarely detected in trophoblastic syncytium. In all abnormal placentae, the LHRI of sinusoids was absent, and syncytium showed either enhanced (immature placental phenotypes) or no LHRI (aged placental phenotype). LHRI in human brain was identified in microglial cells (CD68+ resident macrophages). Protein extracts from human vaginal wall and levator ani muscle and fascia showed strong approximately 92 and 68 kDa species, and LHRI was detected in smooth muscle cells, fibroblasts, resident macrophages and nuclei of skeletal muscle fibers. Our observations indicate that, in contrast to the theory on the role of vascular hormone receptors in preferential pick up of circulating hormones, there is no need to enhance selective pick up rather only prevent LH/CG transport to inappropriate sites. Abnormal placental LHR expression may play a role in the development of abnormal pregnancy. Expression of LHR in the pelvic floor compartments suggests that high LH levels in postmenopausal women may contribute to the pelvic floor relaxation and increased incidence of pelvic floor disorders. Since chorionic gonadotropin increases secretion of a variety of cytokines by monocytes, and induces their inflammatory reaction and phagocytic activity, high LH levels in aging individuals may also activate microglia (mononuclear phagocyte system in the central nervous system) and contribute to the development of Alzheimer's disease and other inflammation-mediated neurodegenerative diseases.

Show MeSH
Related in: MedlinePlus