Exogenous long-term treatment with 17ß-oestradiol alters the innervation pattern in pig ovary.

The aim of the present study was to determine the effect of long-term 17ß-oestradiol (E2) exposure, a simulation of pathological states that occur with oestrogen overproduction, on the innervation patterns of ovaries in adult gilts. The intraovarian distribution and density of nerve fibres immunoreactive (IR) to protein gene product (PGP) 9.5 and containing dopamine-ß-hydroxylase (DBH), neuropeptide Y (NPY), somatostatin (SOM) and galanin (GAL) were determined. From Day 4 of the first oestrous cycle to Day 20 of the second cycle studied, experimental gilts were injected with E2 (1000µg every 12h) whereas control gilts were injected with corn oil. After E2 administration, there was an increase in the number of PGP9.5-, DBH-, NPY- and GAL-IR fibres. Numerous PGP9.5-IR terminals were observed within the ground plexus around secondary follicles and small or medium tertiary follicles. Long-term E2 treatment increased the density of DBH- and NPY-IR fibres in the cortical part of the ground plexus, DBH- and GAL-IR fibres in the medullary part of the ground plexus, DBH-IR fibres near small and medium tertiary follicles and NPY-IR fibres around medullary arteries. The data indicate that long-term exposure of gilts to E2 increases the total number of intraovarian fibres, including sympathetic fibres. These results suggest that elevated E2 levels that occur during pathological states may affect the innervation patterns of ovaries and their function(s).

Long-term estradiol-17ß administration reduces population of neurons in the sympathetic chain ganglia supplying the ovary in adult gilts.

Elevated levels of endogenous estrogens occurring in the course of pathological states of ovaries (follicular cysts, tumors) as well as xenoestrogens may result in hyperestrogenism. In rat, a close relationship between estrogens and sympathetic and sensory neurons supplying the genito-urinary system was reported. Recently, we have shown that long-term estradiol-17ß (E(2)) administration affected morphological and immunochemical organization of the sympathetic ovarian neurons in the caudal mesenteric ganglion of adult gilts. In this study, the influence of E(2) overdose on the number and distribution of neurons in the sympathetic chain ganglia (SChG) projecting to the ovary of adult pigs was investigated. The numbers of ovarian dopamine-ß-hydroxylase (DßH-), neuropeptide Y (NPY-), somatostatin (SOM-), galanin (GAL-) and estrogen receptors (ERs-) immunoreactive perikarya as well as the density of the intraganglionic nerve fibers containing DßH and/or NPY, SOM, GAL were also determined. On day 3 of the estrous cycle the ovaries of both the control and experimental gilts were injected with retrograde neuronal tracer Fast Blue, to identify the neurons innervating gonads. From day 4 of the estrous cycle to the expected day 20 of the second studied cycle, the experimental gilts were injected with E(2), while the control gilts were receiving oil. After the last E(2)/oil injection, the SChG Th16-S2 were collected and processed for double-labeling immunofluorescence. Injections of E(2): (1) increased the E(2) level in the peripheral blood ~4-5 fold, (2) reduced the total number of Fast Blue-positive postganglionic neurons in the ganglia under investigation, (3) decreased the number of perikarya in the L2-L4 ganglia, (4) reduced the number of perikarya in the ventral, dorsal and central regions of the SChG, (5) decreased the numbers of DßH(+)/NPY(+) and DßH(+)/GAL(+) perikarya and the numbers of DßH(+) but NPY(-), SOM(-) and GAL(-) perikarya in the SChG, (6) decreased the number of perikarya expressing ERs subtype α and ß, and (7) decreased the total number of the intraganglionic nerve fibers containing DßH and/or NPY. These results show that long-term E(2) treatment of adult gilts down-regulates the population of both noradrenergic and ERs expressing the SChG ovary supplying neurons. Our findings suggest also that elevated E(2) levels that occur during pathological states may regulate gonadal function(s) by affecting ovary supplying neurons.

Expression of nerve growth factor and its receptors, TrkA and p75, in porcine ovaries.

The cellular localization of nerve growth factor (NGF) and its receptors (TrkA, p75) was investigated during the estrous cycle in gilts. Also, the levels of expression of these factors in walls of tertiary follicles and corpora lutea (CLs) were determined using Western blot. The ovaries from days 3, 7, 16 and 20 of the cycle revealed the presence of NGF and its receptors in oocytes of secondary and tertiary follicles, follicular cells of primary and secondary follicles, thecal and granulosa cells of tertiary follicles and steroidogenic cells of CLs. In wall cells of primary follicles, NGF, TrkA and p75 staining was strongest on day 16, while in secondary follicles, only p75 was more intensely stained on day 16 and 20. In walls of small (to 3 mm in diameter) and medium (4-6 mm in diameter) follicles, NGF staining was lower on day 16, and the p75 reaction was strongest on day 20. On day 20, NGF staining in large follicles (7-10 mm in diameter) was higher than in smaller follicles. The levels of NGF and p75 in small and medium follicles were highest on day 20. The contents of NGF and TrkA in large follicles on day 20 were higher than in smaller follicles. NGF and TrkA contents in CLs were highest on day 7. Our study demonstrates that NGF, TrkA and p75 are expressed in the ovary during the estrous cycle in gilts. These results suggest that NGF and its receptors may be important for ovarian function in cycling gilts.

Long-term estradiol-17ß administration decreases the number of neurons in the caudal mesenteric ganglion innervating the ovary in sexually mature gilts.

The effect of estradiol-17ß (E(2)) on the number and distribution of neurons in the caudal mesenteric ganglion (CaMG) supplying the ovary of adult pigs was investigated. Also, the numbers of ovarian dopamine-ß-hydroxylase (DßH-), neuropeptide Y (NPY-), somatostatin (SOM-), galanin (GAL-) and estrogen receptor (ER)-immunoreactive perikarya as well as the density of the intraganglionic nerve fibers containing DßH and/or NPY, SOM, GAL were determined. E(2) was administered i.m. from day 4 of the first studied estrous cycle to the expected day 20 of the second studied cycle. Injections of E(2) (1) increased the E(2) level in the peripheral blood approximately 4-5 fold, (2) decreased the number of small-sized Fast Blue-positive postganglionic neurons in the CaMG, (3) decreased the number of small perikarya in the ventral, dorsal and central regions of the CaMG, (4) decreased the number of large perikarya in the dorsal and central regions, (5) decreased the number of small and large perikarya in the CaMG that were DßH(+)/NPY(+), (6) decreased the number of small DßH(+) but NPY(-) perikarya, (7) decreased the number of small perikarya coded DßH(+)/SOM(+) and DßH(+)/SOM(-), (8) decreased the number of small DßH(+)/GAL(-) perikarya, (9) decreased the number of small and large perikarya expressing ER subtypes α and ß and (10) decreased the total number of nerve fibers in the CaMG containing DßH and/or NPY and DßH and/or GAL. These results show that long-term E(2) treatment of adult gilts downregulates the populations of both noradrenergic and ERs expressing ovarian neurons in the CaMG. Our findings suggest also that elevated E(2) levels that occur during pathological states may regulate gonadal function(s) by affecting ovary supplying neurons.

Distribution and chemical coding of sympathetic neurons in the caudal mesenteric ganglion projecting to the ovary in sexually mature gilts.

The distribution and co-localisation patterns of dopamine-beta-hydroxylase (DßH), neuropeptide Y (NPY), somatostatin (SOM) and galanin (GAL) were investigated by use of retrograde neuronal tracing and double-labelling immunofluorescence techniques in the caudal mesenteric ganglion (CaMG) neurons supplying the ovary of adult pigs. The existence and density of nerve fibres that are immunoreactive (IR) for the above-mentioned neuroactive substances were also evaluated. Injections of a fluorescent tracer (Fast Blue; FB) into the ovaries revealed the presence of small- (76.38%) and large-sized (23.62%) FB-positive postganglionic neurons in the CaMG. Noradrenergic FB-positive cells were simultaneously NPY- (43.38%), SOM- (18.77%) and GAL- (18.31%) IR. Of the examined FB-positive neurons, 53.49% were DßH-IR but NPY-immunonegative (IN), 79.06% were DßH-IR but SOM-IN, and 77.16% were DßH-IR but GAL-IN. Small- or large-sized subsets of traced neurons were supplied by only one or a few nerve fibres, exhibiting DßH-, NPY-, SOM- and/or GAL-IR. Our data show the specific morphological as well as immunochemical structural organisation of the sympathetic neurons in the CaMG in adult gilts. The occurrence of an abundant population of noradrenergic perikarya in the CaMG may suggest their important physiological role in the regulation of gonadal function(s) in these animals.