Rajani Choudhuri

Determination of the number and distribution of macrophages, lymphocytes, and granulocytes in the mouse uterus from mating through implantation.

The concentration and distribution of F4‐80 positive cells (macrophages) and common leukocyte antigen (CLA) positive (bone marrow derived) cells were assessed in mouse uterus between days 1 and 8 of pregnancy. High numbers of polymorphonuclear leukocytes and lymphocytes were present on days 1 and 2, but not thereafter. Granulocytes were found both in the endometrium and within the luminal epithelium. The percentage of total cells contributed by macrophages was high on days 1 and 2. That percentage decreased significantly on day 3, then increased again on day 5 and remained high through day 8. Macrophages always were found in myometrial stroma. Macrophages were found throughout the endometrium on days 2 through 8. High numbers of macrophages were observed near epithelia, particularly on days 1,2,4, and 5. Few F4‐80+ or CLA+ cells were observed within the developing primary and secondary decidua. The results demonstrate that an inflammation‐like cellular response occurs in the uterus following mating and that macrophages are a major cellular component of the uterus during early pregnancy.

Relative role of CSF‐1, MCP‐1/JE, and RANTES in macrophage recruitment during successful pregnancy

It has been previously demonstrated that macrophage colony stimulating factor (CSF‐1) is produced by uterine epithelial cells in response to estrogen and progesterone. Studies in normal and op/op mice demonstrated that accumulation of a portion of the uterine macrophage population could be attributed to the chemotactic properties of CSF‐1. Op/op mice exhibit greatly reduced rates of fertility, but successful pregnancy is not completely blocked. Also, uteri from op/op mice are not completely macrophage deficient. There are two possible explanations for this. One is that not all tissue macrophages are recruited from the bone marrow pool; some may be derived from primitive mesenchyme. Alternatively, tissue macrophages may be recruited from the bone marrow pool through expression of other type I chemokines such as JE, RANTES, MIP‐1α, MIP‐1β, IP‐10, and KC. Both RANTES and JE are expressed at higher levels than CSF‐1 during early pregnancy. The variable expression and relative role of these various chemokines in pregnancy was addressed by measuring mRNA expression during the first 8 days of pregnancy and in a pseudopregnant model. The expression of these various genes relative to macrophage numbers and macrophage distribution will be discussed. The relative role of these various factors in preparing the uterus for blastocyst implantation will be discussed. Mol Reprod Dev 46:62–70, 1997.

Cortical spreading depression and gene regulation: Relevance to migraine

Cortical spreading depression (CSD) may be the underlying mechanism of migraine aura. The role of CSD in initiating a migraine headache remains to be determined, but it might involve specific changes in gene expression in the brain. To examine these changes, four episodes of CSD at 5‐minute intervals were induced in the mouse brain by application of 300mM KCl, and gene expression was examined 2 hours later using cDNA array and reverse transcriptase–polymerase chain reaction. Controls consisted of groups that received anesthesia only, attachment of recording electrodes only, and application of 0.9% NaCl. Of the over 1,180 genes examined in our experiments, those consistently regulated by CSD included vasoactive peptides; the vasodilator atrial natriuretic peptide was induced by CSD, while the vasoconstrictor neuropeptide Y was downregulated. Other genes specifically regulated by CSD were involved in oxidative stress responses (major prion protein, glutathione‐S‐transferase‐5, and apolipoprotein E). L‐type calcium channel mRNA was upregulated. In summary, CSD regulates genes that are intrinsic to its propagation, that identify accompanying vascular responses as a potential source of pain, and that protect against its potential pathological consequences. We believe these observations have strong relevance to the mechanisms of migraine and its outcomes.

Macrophage colony stimulating factor controls macrophage recruitment to the cycling mouse uterus.

Progesterone and estrogen induce uterine epithelial cells to produce macrophage colony stimulating factor (CSF-1), and mouse uterine epithelial cells produce large amounts of CSF-1 during embryo development. The present study demonstrated that estrogen and progesterone each induce uterine CSF-1 gene transcription and translation detectable by Northern blotting and bioassay. Intrauterine CSF-1 production was greater in the presence of both estrogen and progesterone than following exposure to either hormone alone. CSF-1 mRNA was detectable in the uterus throughout the estrous cycle while CSF-1 bioactivity was detected only during proestrus. CSF-1 production was directly related to the presence of macrophages in the uterus. Ovariectomy, which was rapidly followed by a loss of uterine CSF-1 gene transcription, also was followed by a dramatic decrease in the number of uterine macrophages. Both uterine CSF-1 and uterine macrophages were reconstituted in a dose-dependent manner by systemic administration of estrogen or progesterone. High concentrations of circulating estrogen and progesterone increased the number of macrophages in the uterus and increased their accumulation near epithelial surfaces. Similar relationships were observed in the uterus of cycling mice. Macrophages accumulated in the uterus following intraluminal injection of recombinant human CSF-1 to ovariectomized mice, directly demonstrating the ability of CSF-1 to recruit macrophages from peripheral blood into the uterus. These studies demonstrated that estrogen and progesterone stimulation of CSF-1 production by mouse uterine epithelial cells controls recruitment and distribution of macrophages in the uterus during the estrous cycle.

Early dysregulation of cripto-1 and immunomodulatory genes in the cerebral cortex in a macaque model of neuroAIDS

Human immunodeficiency virus type 1 (HIV-1) and related primate lentiviruses are known to enter the central nervous system (CNS) during the primary phase of infection. Neuroinvasion by simian immunodeficiency virus and simian human immunodeficiency virus (SHIV) is characterized by transient meningitis and astrocytosis. In this report, we used targeted cytokine cDNA arrays to analyze cortical brain tissue from four pig-tailed macaques inoculated for 2 weeks with pathogenic SHIV(50OLNV) and a normal age-matched pig-tailed macaque. Our results revealed that eight genes were significantly upregulated in all four macaques. These included: leukocyte interferon inducible peptide, corticotrophin releasing factor receptor 1, interleukin 6, CDW40 antigen, cysteine-rich fibroblast growth factor, neurotrophin 3, ciliary neurotrophin factor receptor and cripto-1. The upregulation of three of these genes was confirmed by reverse transcriptase PCR (RT-PCR). Since cripto-1 had not been previously identified within specific cell types within the primate central nervous system, we performed immunohistochemical studies, which revealed the presence of cripto-1 in neurons. RT-PCR studies demonstrated that cripto-1 mRNA was widely expressed in the CNS. These results indicate that immunomodulatory genes are upregulated during the primary phase of infection of the central nervous system. Cripto-1, which acts as a survival factor in tumor cells and may be neuroprotective, is expressed in neurons within the CNS and is upregulated during viral invasion.

Leukocyte distribution in the pseudopregnant mouse uterus.

ABSTRACT: Pseudopregnancy, induced by mating females with vasectomized males, is a frequently used model for studying pregnancy‐like uterine changes in the absence of an embryo. Leukocytes make a significant contribution to uterine cellularity during pregnancy. The present study was designed to determine whether changes in numbers and distribution of leukocytes in the uterus during pseudopregnancy and following intraluminal injection of a deciduogenic stimulus parallel changes observed during the first eight days of pregnancy. Common leukocyte antigen positive (CLA +) cells, macrophages (F4/80+ cells), and granulocytes were assessed between days 1 and 8 of pseudopregnancy using qualitative and quantitative immunohistochemistry. High numbers of CLA+ leukocytes were present on days 1 and 2. Those were comprised primarily by macrophages, polymorphonuclear leukocytes, and eosinophils. High concentrations of leukocytes were detected in the endometrium, and some granulocytes were observed migrating through the luminal epithelium. Leukocytes, principally macrophages, were reduced in number and were distributed throughout the endometrium on days 3 and 4. Introduction of oil to the uterine lumen on day 4 stimulated primary decidualization. Decidual cells were CLA − and F4/80, − and, as decidualization proceeded, CLA+ and F4/80+ cells decreased in number in the anti‐mesometrial uterus and were detected primarily in the deep endometrium. Later, a secondary decidualization zone developed in the mesometrial aspect of the uterus. Unlike the initial decidual reaction, which was relatively free of leukocytes, the secondary decidual zone contained very high numbers of CLA+ and F4/80+ cells. The uninjected uterine horn remained relatively unchanged from days 3 through 8. The quantitative and distributional changes observed in the leukocyte populations during pseudopregnancy were similar to those which have been observed during pregnancy.