In the preluteolytic animals the most intense signal was seen in a CL collected from a cow (number 959) that, out of the three preluteolytic animals studied, had the highest plasma PGFM concentration and lowest progesterone concentration at the time of tissue collection (Table 1). Buy Advair Diskus Online
Immunohistochemistry was performed on serial sections from the same CL used for in situ hybridization (Fig. 4). Cells that stained positively with the CD5 + T lymphocyte marker were present in all sections of CL studied. They were distributed throughout the luteal tissue as individual cells and in small clumps. Positive staining was absent from negative control samples (Fig. 4c).
When compared to serial sections from the in situ hybridization studies, areas of luteal tissue were seen in which the distribution of cells expressing mRNA for MCP-1 was similar to the distribution of T lymphocytes (Fig. 4, a and b); however, there were also other regions where this was not the case. In addition, significant numbers of T lymphocytes were also present in CL where MCP-1 mRNA expression was minimal (preluteolysis).
FIG. 3. Darkfield illuminations of sections (14 ^m) from CL collected before (a and b) and after (c and d) functional luteolysis. Sections were probed with 35S-labeled homologous antisense (a-c) and sense (d) bovine MCP-1 RNA probes (X240).
FIG. 4. Serial sections of a CL collected after functional luteolysis viewed under darkfield illumination (a) after probing with 35S-labeled homologous antisense bovine MCP-1 RNA and lightfield illumination (b) after immunohistochemical staining with an antibody specific for CD5 + T lymphocytes; (c) immunohistochemistry negative control. X240 (reproduced at 87%).