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PubMed1993

The origin of cytologically unidentifiable chromosome abnormalities: six cases ascertained by targeted chromosome-band painting.

De novo chromosome structural abnormalities cannot always be diagnosed by the use of standard cytogenetic techniques. We applied a previously developed chromosome-band-specific painting method to the diagnosis of such rearrangements. The diagnostic procedures consisted of microdissection of an aberrant chromosomal region of a given patient, polymerase chain reaction (PCR) amplification of the dissected chromosomal DNA, and subsequent competitive fluorescence in situ hybridization (FISH) using the PCR products as a probe pool on metaphase chromosomes from the patient and/or a karyotypically normal person. With this strategy, we studied 6 de novo rearrangements (6p+, 6q+, 9p+, 17p+, +mar, and +mar) in 6 patients. These rearrangements had been seen by conventional banding but their origin could not be identified. In all 6 patients, we successfully ascertained the origin. Using an aberrant region-specific probe pool, FISH signals appeared on both the aberrant region and a region of another specific chromosome pair. A reverse probe pool that was generated through the microdissection of normal chromosomes at a candidate region for the origin of the aberration hybridized with both the aberrant and the candidate regions. We thus diagnosed one patient with 17p+ as having trisomy for 14q32-qter, one with 9p+ as having trisomy for 12pter-p12, one with 6q+ as having a tandem duplication (trisomy) of a 6q23-q25 segment, one with 6p+ as having a tandem duplication (trisomy) of a 6p23-q21.3 segment, one with a supernumerary metacentric marker chromosome as having tetrasomy for 18pter-cen, and the last with an additional small marker chromosome as having trisomy for 18p11.1 (or p11.2)-q11.2. The present targeted chromosome-band-painting method provides the simple and rapid preparation of a probe pool for region-specific FISH, and is useful for the diagnosis of chromosome abnormalities of unknown origin.

Base Sequence↗
PubMed1991

Mating alters topography and content of oxytocin immunoreactivity in male mouse brain.

Sexual stimulation of males has been reported to affect hypothalamic oxytocinergic systems. In the present study we used radioimmunoassays of microdissected forebrain regions and immunocytochemical analysis of Vibratome sections to study the oxytocin systems of naive males, males killed after one mating, and males mated daily with different receptive females for 3 weeks. In males that had mated once, less oxytocin-immunoreactive neurons were observed in the paraventricular (PVN), supraoptic (SON) and periventricular (NPE) nuclei than in naive males. However, after repeated matings, the number of immunoreactive neurons and their staining intensity was increased in these regions. Furthermore, additional oxytocinergic neurons could be found in the lateral subcommissural nucleus, the zona incerta and the ansa lenticularis of repeatedly mated males. Oxytocin-immunoreactive neurons were only occasionally seen in these areas in unmated males or in animals that had been killed after initial mating. Radioimmunoassays of microdissected PVN, SON, NPE and the lateral hypothalamus confirmed the reduction in oxytocin-immunoreactive levels after a first mating by a male and the increase after repeated matings. It is likely that oxytocin secretion into peripheral and portal circulation is stimulated by the endocrine conditions associated with initial mating. These immediate effects may be followed by the activation of synthesis in oxytocin neurons in several sites of the basal forebrain.

Animals↗
PubMed1987

Roentgenological investigation of cavernous sinus structure with special reference to paracavernous cranial nerves.

We have evaluated the anatomical structure of cavernous sinuses in parasellar blocks obtained from adult cadavers to gain clinically useful information. Ten microdissectional studies of parasellar blocks (20 sides of cavernous sinus) were carried out with special emphasis on the course of paracavernous cranial nerves (3rd to 6th). These were also examined on plain radiographs. Secondly, CT investigations of the cavernous sinuses (notably as to paracavernous cranial nerves) were undertaken in twenty clinical cases having presumed parasellar lesions and in three parasellar blocks from cadavers. The results from microdissectional studies and plain radiographs were as follows. It was found that the 3rd cranial nerve entered the lateral wall of the cavernous sinus close to the antero-inferior part of the posterior clinoid on lateral projection. The 4th cranial nerve, on the other hand, entered near the postero-inferior portion of the posterior clinoid on the same view. Variations in Parkinson's triangle were fairly often noted, although marked asymmetry was not seen in individual cases. From CT findings, it was found that a postenhanced, magnified direct coronal study was essential for identifying paracavernous cranial nerves more clearly. The 3rd and 5th cranial nerves were fairly well identified on axial and coronal projections. Careful radiological investigation including plain skull radiographs & CT seems to be of value for those who diagnose or treat parasellar lesions.

Abducens Nerve↗
PubMed1989

Antigenic expression of aminopeptidase M, dipeptidyl-peptidase IV and endopeptidase by primary cultures from rabbit kidney proximal tubule.

Techniques using microdissected tubules from rabbit kidney allow the isolation of well defined segments which can be cultured to obtain pure renal cell epithelia. From microdissected proximal tubules, we obtained epithelia the cells of which exhibit some of the antigenic expressions of the initial proximal cells. For this purpose, we used three monoclonal antibodies raised against apical brush border membranes of the proximal tubules. We determined with precision the identity and some of the molecular characteristics of the antigens bound by these three antibodies and found that they correspond to three hydrolases present in the brush borders of proximal renal cells (amino-peptidase, dipeptidyl-peptidase IV and endopeptidase). These apical markers are expressed by the growing cells of primary cultures from proximal tubules, suggesting strongly that they are effectively proximal cells and that no appreciable dedifferentiation occurred during the growth process. We have also shown that apical expression of these hydrolases on the plasma membrane of the epithelium occurred only after several days of culture and determined the complete polarization of the cells. Electron microscopy studies confirmed the degree of polarization of the cultured cells by the presence of numerous microvilli on their apical face.

Aminopeptidases↗
PubMed1975

Evaluation of peritubular capillary microperfusion method by morphological and functional studies.

The present study was undertaken in anesthetized rats to evaluate the technique of efferent arteriolar and branch capillary microperfusion in the kidney. In contrary to previous suggestions, subsurface efferent arteriolar branches were demonstrated to occur prior to the surface site of insertion of the microperfusion pipette into an efferent arteriole. This finding, in addition to the presence of continued blood flow around the microperfusion pipette, provided an explanation for continued filtration and tubular reabsorption in nephrons under study. Moreover, additional microdissection studies demonstrated that a mean of 3.8 nephrons are surrounded by artificially perfused capillaries from a micropipette inserted into a single efferent arteriole. Thus, any obstructive effect of the microperfusion technique on post-glomerular blood flow might be expected to alter nephron function in only a minority of nephrons which are surrounded by artificial perfusate. The results also demonstrated the importance of avioding osmotic gradients between the animal's plasma and the artificial perfusate. A method is also described in which the correlation between in vivo diagrams and in vitro microdissections allows assessment of the completeness of the artificial perfusion of capillaries surrounding the surface loops of nephrons under study.

Animals↗
PubMed1997

Loss of heterozygosity of the retinoblastoma gene in liver cirrhosis accompanying hepatocellular carcinoma.

Carcinogenesis is a multistep process. Most hepatocellular carcinoma (HCC) is preceded by liver cirrhosis, but the genetic changes involved in cirrhosis are not known well. The present study was conducted to evaluate aberration of the retinoblastoma (RB) gene in HCC and adjacent non-tumorous liver using 22 patients with chronic liver damage accompanying HCC. The specimens obtained by microdissection from paraffin-embedded tissues were analyzed using an assay based on the polymerase chain reaction for highly polymorphic nucleotide sequences of microsatellites in the RB gene. Out of 22 cases, 15 showed constitutional heterozygosity for the microsatellite markers. In 11 (73.3%) of these 15 informative cases, the primary HCC foci showed loss of heterozygosity (LOH). In 8 of these 11 doubly informative (informative and LOH-positive in primary HCC) cases, LOH was found in 20 (64.5%) of 31 microdissected non-tumorous foci. All of the non-tumorous foci showing RB loss were cirrhotic lesions but there were no foci of chronic hepatitis. The remaining 4 cases without LOH in HCC foci showed no LOH in non-tumorous lesions. In our study, LOH of the RB gene was frequently observed in liver cirrhosis surrounding tumor.

Aged↗
PubMed1984

Comparison of tritiated thymidine and metaphase arrest techniques of measuring cell production in rat intestine.

Measurements of intestinal cell production at several sites throughout the gastrointestinal tract were compared using two different methods, namely tritiated thymidine to determine the uptake of tritium per unit weight of tissue (dpm/mg) and per microdissected intestinal crypt or stomach gland (dpm/crypt or gland) and the metaphase arrest technique, in the same group of rats. The metaphase arrest technique was employed to determine the crypt cell production rate per hour (CCPR). The dpm/crypt or gland of tritiated thymidine-derived tritium in microdissected intestinal crypts or stomach glands showed a good linear correlation (r = 0.93) with CCPR which extrapolated through the origin. The dpm/mg wet weight of tissue showed a good linear correlation with CCPR (r = 0.87) and dpm/crypt (r = 0.97); however, neither of these relationships extrapolated through the origin. Instead there was a positive intercept of dpm/mg tissue which represented about 25-30% of the maximal tritium content of the intestine. This radioactivity in tissue had no counterpart in the crypts or glands and presumably reflected unbound tritium and tritium in cells which were located outside the epithelial compartment. The good linear correlation (r = 0.93) between dpm/crypt and CCPR extended to the intestine of transected and 75% small bowel resected rats. Subsequent analysis of the times recorded for these procedures showed that determination of CCPR was faster by at least 25% than measurement of dpm/crypt. In conclusion, dpm/crypt but not dpm/mg tissue gave a valid estimate of intestinal cell production equivalent to measurement of CCPR.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1977

Isolation of viable human pancreatic islets.

This article reports a method for the isolation of viable pancreatic islets from the human pancreas. Isolated islets were obtained from human pancreata of cadavers, patients undergoing surgical operations, and fetuses, using a freehand microdissection procedure. Viability was assessed by light microscopy of sections stained with aldehyde fuchsin and by measuring the insulin output of islets in response to a glucose stimulus in vitro using a perifusion system. Ten pieces of cadaver pancreas were studied. Islets were isolated from 6 specimens and in 5 of these were shown to respond to a glucose stimulus in vitro. Histologically the islets showed minimal damage with slight degranulation of the beta cells. Five pieces of pancreas removed at operation were studied as well. Islets were isolated in all cases, but only 2 showed a response to a glucose stimulus. Pancreata from a 26-week and 34-week human fetus were also studied. It was not possible to microdissect islets from either case, but small pieces of pancreas from the 26-week fetus were shown to respond to a glucose stimulus by producing a significant increase in insulin output.

Cadaver↗
PubMed1982

Structural and functional correlations in the hypertrophic mucosa of intestinal remnants following resection in rats.

Microdissection techniques were applied to determine the absolute increase in villus size and absorptive surface in the mucosa of rat intestinal remnants 3 weeks after 60% proximal resection. Adjacent tissues were used to evaluate the operational kinetic parameters describing the influx of L-phenylalanine and alpha-methyl-D-glucoside into intestinal rings. The villi of ileal remnants were enlarged significantly in all dimensions; the mean surface area of the villus was 2.5 times, and mucosal:serosal surface ratio 1.9 times larger than normal. The influx of phenylalanine and alpha-methyl-glucoside into the mucosa of ileal remnants was reduced, but the change could not be ascribed to an attenuation of one specific kinetic parameter. Using the microdissection data to transform the Vmax-values to a mucosal area basis, we could demonstrate that the Vmax for both substrates was smaller than normal. The results may derive from changes in the maturity of the cell population in intestinal remnants.

Animals↗
PubMed2006

Beta cell death in hyperglycaemic Psammomys obesus is not cytokine-mediated.

AIMS/HYPOTHESIS: It has recently been proposed that IL-1beta may be responsible for beta cell death in type 2 diabetes mellitus. Major support for this assumption was derived from experiments in the gerbil Psammomys obesus (sand rat), a model for nutritionally induced non-insulin-dependent type 2 diabetes. Using gerbil-specific primers for the analysis of gene expression, we investigated the validity of this hypothesis. METHODS: Gene expression was analysed by real-time RT-PCR of isolated and laser-microdissected islets and by in situ RT-PCR, both in beta cells and in immune cells, as well as in lymph nodes and spleen. RESULTS: We were unable to detect Il-1beta and the IL-1beta-inducible enzyme inducible nitric oxide synthase (iNos) by in situ RT-PCR, either in the pancreatic beta cells, or in the small number of non-activated immune cells of healthy and diabetic Psammomys obesus after 1 and 3 weeks on a high-energy diet. Very low levels of Il-1beta and iNos mRNA were detectable in collagenase-isolated and laser-microdissected islets of normoglycaemic gerbils by real-time RT-PCR without any increase of these mRNAs in islets from diabetic animals. These results were confirmed by electron microscopy with immunogold staining for IL-1beta and insulin. CONCLUSIONS/INTERPRETATION: The diabetic syndrome induced in Psammomys obesus by high-energy diet is a classical type 2 diabetes model, which does not show any evidence of an involvement of the proinflammatory cytokine IL-1beta or of activated immune cells in its pathogenesis. This is clearly at variance with the situation in type 1 diabetes.

Animals↗
PubMed2004

Distribution of HLA class I altered phenotypes in colorectal carcinomas: high frequency of HLA haplotype loss associated with loss of heterozygosity in chromosome region 6p21.

HLA class I loss or down-regulation is a widespread mechanism used by tumor cells to avoid tumor recognition by cytotoxic T lymphocytes, and thus favor tumor immune escape. Multiple mechanisms are responsible for these HLA class I alterations. In different epithelial tumors, loss of heterozygosity (LOH) at chromosome region 6p21.3, leading to HLA haplotype loss, occurs in 6-50% of all cases depending on the tumor entity. In this paper we report the frequency of LOH at 6p21 in 95 colorectal carcinomas (CRC) previously analyzed for altered HLA class I expression with immunohistological techniques. We used PCR microsatellite amplification of selected STR markers located on Chromosome 6 to identify LOH with DNA from microdissected tumor tissues and the surrounding stroma. Sequence-specific oligonucleotide analysis was performed in microdissected stroma and tumor cells for HLA typing, and to detect HLA haplotype loss. A high frequency (40%) of HLA haplotype loss was found in CRC. Eight tumors showed microsatellite instability. We sometimes observed two or more mechanisms responsible for HLA alteration within the same HLA-altered phenotype, such as LOH and HLA class I total loss. In 25 tumors (26%) no HLA class I alteration could be identified. These data are potentially relevant for CRC patients undergoing T-cell-based immunotherapy.

Chromosomes, Human, Pair 6↗
PubMed2002

Multiple mechanisms generate HLA class I altered phenotypes in laryngeal carcinomas: high frequency of HLA haplotype loss associated with loss of heterozygosity in chromosome region 6p21.

Major histocompatibility complex (MHC) class I loss or downregulation in cancer cells is a major immune escape route used by a large variety of human tumors to evade anti-tumor immune responses mediated by cytotoxic T lymphocytes. Multiple mechanisms are responsible for such HLA class I alterations. However, the precise frequency of these molecular defects has not been clearly determined in tumors derived from specific tissues. To analyze such defects we aim to define the major HLA class I-altered phenotypes in different tumor types. In this paper we report on HLA class I expression in 70 laryngeal carcinomas. We used immunohistological techniques with a highly selective panel of anti-HLA monoclonal antibodies (mAb), and polymerase chain reaction (PCR) microsatellite amplification of previously selected microsatellite markers (STR) located in chromosome 6 and 15. DNA was obtained from microdissected tumor tissues and surrounding stroma to define the loss of heterozygosity (LOH) associated with chromosome 6p21. Our results showed that LOH in chromosome 6 produced HLA haplotype loss (phenotype II) in 36% of the tumors. In addition, HLA class I total loss (phenotype I) was found in 11%; HLA A or B locus downregulation (phenotype III) was detected in 20%; and HLA class I allelic loss (phenotype IV) in 10% of all cases. We sometimes observed two or more associated mechanisms in the same HLA-altered phenotype, such as LOH and HLA total loss in phenotype I. In only 23% of tumors it was not possible to identify any HLA class I alteration. We conclude that the combination of immunohistological techniques and molecular analysis of tumor DNA obtained from microdissected tumor tissues provides a means for the first time of determining the actual frequency of the major HLA class I-altered phenotypes in laryngeal carcinomas.

Antigens, Neoplasm↗
PubMed1998

Determinative factors of biochemical cure after primary and reoperative surgery for sporadic medullary thyroid carcinoma.

Normalization of calcitonin levels after surgery has been regarded as the most powerful prognostic factor for medullary thyroid carcinoma (MTC). Although the prognosis of patients with persistent hypercalcitoninemia may be acceptable, the biochemical cure rate can be improved by new microdissection techniques. This raises certain questions: Can extension of locoregional lymphadenectomy (LA) further improve biochemical cure and survival after primary or reoperative MTC surgery? Which factors concerning TNM categories are associated with the possibility of postoperative normalization of calcitonin levels? This study included 64 patients with sporadic MTC operated on from 1986 to 1997. Altogether 27 patients underwent primary surgery, and 37 patients were reoperated, performing a microdissection of all four locoregional compartments (four-compartment lymphadenectomy, or 4CLA). For primary MTC the biochemical cure rate was 100% in node-negative patients and 33% in node-positive patients; the latter could be improved to 45% after 4CLA. In contrast to reoperative MTC, the rate of lymph node metastases (LNMs) with primary MTC correlated with the pT category (pT1 33%, pT2 53%, pT3 100%, pT4 100%) but not with age or sex. Again in contrast to reoperative MTC, mediastinal LNMs in primary MTC were present only in patients with a pT4 tumor. At reoperation, 4CLA was able to cure 22% of node-positive patients, 28% without proved distant metastases. No patient with extrathyroidal tumor involvement or distant metastases was biochemically cured after either primary or reoperative surgery. For all node-positive MTC patients, in addition to cervicocentral LA at least a bilateral cervicolateral LA is recommended. Transsternal mediastinal lymph node dissection is indicated in patients with LNMs in the cervicomediastinal transition, facilitating biochemical cure in up to 45% after the first operation and 22% after reoperative surgery of sporadic MTC.

Adult↗
PubMed2002

The petromastoid canal on computed tomography.

The objective was to assess visibility and anatomy of the petromastoid canal in high-resolution CT. Computed tomography images of 188 patients were reviewed for delineation of the petromastoid canal. This bony canal connects the mastoid antrum with the cranial cavity and houses the subarcuate artery and vein. The diameter, obtained in the middle portion of the canal, was compared with the diameter of the vestibular and cochlear aqueduct in all patients, and absolute values measured in 20 cases. Collimation was 1 mm in 164 and 2 mm in 24 examinations. Additionally, temporal bone of a cadaver was imaged and microdissected. The petromastoid canal was identified bilaterally in all 164 scans that were obtained with a slice thickness of 1 mm. In 5 of the 24 patients imaged with a collimation of 2 mm, the canal was not visible, most probably due to partial-volume effects. The petromastoid canal had the same diameter as the cochlear aqueduct in 42/44 (right/left), exceeded it in 66/61 and was smaller in 75/78 cases. In comparison to the vestibular aqueduct it had an equal diameter in 38/41 (right/left), exceeded it in 63/61, and was rated as smaller in 82/81 temporal bones. Diameters for the canals were: petromastoid canal 0.51+/-0.04 mm; cochlear aqueduct 0.57+/-0.03; and vestibular aqueduct 0.63+/-0.06 mm. Microdissection of the specimen revealed the entire course of the canal and demonstrated a similar appearance of the structure as in the images. The petromastoid canal can easily be identified on high-resolution, thin-slice CT images. Knowledge of the anatomy of this bony canal prevents misinterpretation as pathological structure, such as fracture line, which might occur if this structure is not known.

Adult↗
PubMed1997

Comparative chromosome painting between two marsupials: origins of an XX/XY1Y2 sex chromosome system.

Cross-species chromosome painting was used to investigate genome rearrangements between tammar wallaby Macropus eugenii (2n = 16) and the swamp wallaby Wallabia bicolor (2n = 10female symbol/11male symbol), which diverged about 6 million years ago. The swamp wallaby has an XX female:XY1Y2 male sex chromosome system thought to have resulted from a fusion between an autosome and the small original X, not involving the Y. Thus, the small Y1 should represent the original Y and the large Y2 the original autosome. DNA paints were prepared from flow-sorted and microdissected chromosomes from the tammar wallaby. Painting swamp wallaby spreads with each tammar chromosome-specific probe gave extremely strong and clear signals in single-, two-, and three-color FISH. These showed that two tammar wallaby autosomes are represented unchanged in the swamp wallaby, two are represented by different centric fusions, and one by a tandem fusion to make the very long arms of swamp wallaby Chromosome (Chr) 1. The large swamp wallaby X comprises the tammar X as its short arm, and a tandemly fused 7 and 2 as the long arm. The acrocentric swamp wallaby Y2 is a 2/7 fusion, homologous with the long arm of the X. The small swamp wallaby Y1 is confirmed as the original Y by its painting with the tammar Y. However, the presence of sequences shared between the microdissected tammar Xp and Y on the swamp wallaby Y2 implies that the formation of the compound sex chromosomes involved addition of autosome(s) to both the original X and Y. We propose that this involved fusion with an ancient pseudoautosomal region followed by fission proximal to this shared region.

Animals↗
PubMed1997

Targeted development of microsatellite markers from the defined region of bovine chromosome 6q21-31.

A methodical strategy for the isolation of microsatellite markers specific for targeted regions of bovine chromosomes is presented. The procedure involves directed microdissection of one defined subchromosomal area, its DOP-PCR-amplification and cloning. With this approach, a library specific to the BTA 6q21-31 chromosomal region was constructed. Eleven unique microsatellite-containing sequences were isolated, converted into sequence-tagged microsatellite sites, and characterized concerning their species-specific origin. Seven primer pairs generated bovine-specific PCR products and provided a set of microsatellite markers that generally revealed high informativity in the HF breed. Linkage analysis assigned six of them to their predefined subchromosomal origin on BTA 6 corresponding to the specific rehybridization signal of the DOP-PCR product generated from the microdissected chromosome area 6q21-31. The results underline the usefulness of the BTA 6q21-31 library for targeted isolation of unique sequences that are specific for the dissected chromosomal region as demonstrated here by the isolation of microsatellite markers.

Animals↗
PubMed2002

Protamine-1 and -2 mRNA in round spermatids is associated with RNA-binding proteins.

RNA-binding proteins in round spermatids have previously been assigned to the coding sequence of Prm1- and Prm2-mRNA. To further characterize this protein-RNA interaction, prior to cDNA synthesis, microdissected cell profiles were digested with different proteases exhibiting a specific cleavage site followed by both conventional and real-time quantitative PCR. Best results were obtained with proteinase K and A followed by factor Xa protease, genenase I, and proteases V8. While enterokinase revealed PCR signals solely for Prm2, no amplification signal was obtained using chymotrypsin. These data suggest a protein segment rich in basic amino acids to be important for the binding to Prm1- and Prm2-mRNA. The fact that phenanthroline treatment instead of protease digestion also resulted in amplification signals suggests the involvement of zinc-finger-like protein-RNA interactions. Employing different primer pairs, RNA-binding proteins were shown to be localized at the 5' end of Prm1- and Prm2-mRNA. Since protein-RNA interactions are a common principle of posttranscriptional regulation of gene expression, the combination of microdissection, protease digestion, and real-time quantitative PCR provides a suitable tool for its investigation in a cell type-specific manner. Furthermore, the presence of RNA-binding proteins within the coding sequence of mRNAs demands proteinase K treatment prior to cDNA synthesis, a compelling necessity for the study of gene expression.

Cell Separation↗
PubMed2005

Cellular localization of estrogen receptor-alpha (ERalpha) and -beta (ERbeta) mRNA in the boar testis.

Boar testes synthesize high amounts of estrogens which are known to stimulate several male sexual functions in a variety of extragonadal target tissues. Possible effects within the testis depend on the existence of the estrogen receptor subtypes alpha and beta (ERalpha, ERbeta). The precise cellular localization of these subtypes within the testis was, so far, based mainly on protein expression studies using different antibodies in several species including boars shows contradictory results. Therefore, we investigated the ERalpha and ERbeta gene expression using RT-PCR of testis homogenates and RT-PCR after UV-single cell microdissection combined with in-situ hybridization of four fertile boars with an average age of 32 weeks. Both ERalpha and ERbeta mRNA were found in testis homogenates. Using in-situ hybridization and UV-single cell microdissection ERalpha mRNA was present in type A and type B spermatogonia up to mid-pachytene primary spermatocytes in stage V-VIII and stage I of the seminiferous epithelial cycle, but not in other cells. ERbeta mRNA was found only in Sertoli cells. Interstitial Leydig cells revealed neither ERalpha nor ERbeta mRNA. The data suggest a direct impact of estrogen in the boar on Sertoli cell function via ERbeta and germ cell formation via ERalpha.

Animals↗