Demonstration of prolactin messenger RNA after amplification in the brain in rats]

By means of immunocytochemistry, a central neuronal network containing a prolactin-like substance has been described in the rat. In order to demonstrate the synthesis of this peptide in these cells, we examined the presence of prolactin messenger RNA (PRL mRNA) in several brain samples including the pituitary gland. Amplification of the PRL mRNA was performed by the polymerase chain reaction technique, followed by southern blotting and hybridization with a specific oligonucleotide. Results showed the presence of the expected cDNA (468 bp) in the hypothalamus. Another cDNA with a lower molecular weight was also observed.     

Quality of a stress-sensitive Cucurbita pepo L. pollen

The quality of Cucurbita pepo L. pollen was studied using field pollinations and the fluorochromatic-reaction test. The extreme sensitivity of this pollen to dehydration and ageing is demonstrated. Controlled stress applied to mature pollen leads to the development of seedless fruits. Molecular signals seem to be involved in the induction of this parthenocarpy. These results indicate the existence of distinct sequences involved in the completion of the fertilization program of pollen. With pollen altered by stress, the fertilization process may be stopped at different stages of its completion. We bring evidence that Cucurbita pepo plants have developed special adaptations in order to compensate for the poor viability of their pollen.Abbreviation FCR fluorochromatic reaction     

In situ mechanical behavior of posterior spinal ligaments in the lumbar region. An in vitro study

The posterior ligaments: ligamentum flavum, articular, interspinous and supraspinous ligaments of twenty five fresh cadaveric intervertebral segments, from T11-T12 to L4-L5, extracted from fourteen spines were tested in tension. A progressive dissection method was used, that is, each segment was tested after first resecting the disk with the ligaments intact and a force-elongation curve obtained. Then one ligament was cut and the test repeated, and so on. The most restrictive ligament was found to be the ligamentum flavum followed by the articular, interspinous, and supraspinous ligaments.     

Viability of Cururbita pepo pollen: biophysical and structural data

During ageing of the short-lived pollen grains of Cucurbita pepo L., water loss was examined in relation to viability using biophysical (¹H-nuclear magnetic resonance, NMR) and cytological methods (fluorochromatic reaction test, freezefracture and scanning electron microscopy). A semi-logarithmic representation of the pollen weight loss demonstrated the complexity of the dehydration process. A the study of proton loss using ¹H-NMR indicated that two major releases water of had taken place, each with different flux rates. Pulse ¹H-NMR experiments showed the occurrene of non-exponential signal decay as a function of time, indicating the existence of different fractions of water in a pollen grain sample. These fractions leave the pollen grain at different times during pollen dehydration, and one of them (that of the so-called vital water) can be related to pollen viability. The quantity of protons giving a signal during pulse ¹H-NMR experiments was very low when the pollen grains were judged to be dead according to the fluorochromatic test. Freeze-fracture replicas of these dead pollen grains (less than 25% water content) showed that the plasma membrane had become detached from the intine surface; this ultrastructural feature might therefore be involved in the loss of pollen viability.Abbreviations A initial amplitude of the NMR signal - A₂ quantity of water charcterized by T_2-2 - A₅ quantity of water characterized by T_2–5 - FCR fluorochromatic reaction - NMR nuclear magnetic resonance - T₂ transverse relaxation time - T_2-2 T₂ measured with 2 ms between each pulse of radiofrequency - T_2–5 T₂ measured with 5 ms between each pulse of radiofrequency     

Hypersensitive Reaction in Petunia

Intercellular fluid (IF) extracted from the apoplastic leaf spaces of Petunia contained up to 6 new soluble proteins, called d proteins (d₀ to d₅). Characterisation of the d proteins has shown that they can be divided into groups and show a genetic variability similar to the PR(b) proteins from Nicotiana. d protein patterns from Petunia interspecific hybrids clearly show that the determinant for the d proteins in sexually transmitted as is the case for b proteins in Nicotiana. d-protein induction in TMV-SM₁ infected Petunia leaves is discussed in relation to the resistance to infection shown by cultivars and hybrids of Petunia.     

Attempts to transform Zea mays via pollen grains

Summary An attempt was made to transfer two sorts of DNA into maize via pollen grains. Controls for both pollen quality and DNA behaviour during the transformation experiments were included. When genomic DNA was used, no transformants were observed among the 1805 seeds screened. With plasmid DNA (harbouring the gene expressing kanamycin resistance in plant cells), three plants with kanamycin resistance were observed among the 1723 seeds screened, although no molecular evidence of transformation was obtained. Experiments indicated that under the conditions used, DNA was being degraded by both pollen and stigma nucleases. Consequently, we attempted to determine protocols which would inhibit these nuclease activities in order to preserve DNA integrity during transformation experiments, thus allowing fertilization. We found that a classic germination medium supplemented by 300 or 600 mM KNO₃, or 20% PEG₁₅₅₀ satisfied all these conditions.Abbreviations BK Brewbaker and Kwack medium - BKS15 Brewbaker and Kwack medium containing 15% (w/v) sucrose - EDTA ethyldiaminetetraacetic acid - FCR fluorochromatic reaction - FP fertilization percentage - PEG polyethylene-glycol     

The spatial association of the sperm cells and vegetative nucleus in the pollen grain ofBrassica

Summary In mature viable pollen ofBrassica oleracea, the pair of sperm cells and the nucleus of the vegetative cell are linked to form a structured unit we term the male germ unit. The sperm cells are held within a common periplasm and have no cell walls. Each sperm cell has a central globular body containing the nucleus surrounded by several evaginations which provide the means for linkage between them. One sperm cell, usually that closest to the nucleus of the vegetative cell contains most of mitochondria profiles (plastids are absent). This sperm cell appears to be linked by its protoplasmic evaginations to the envelope of the vegetative nucleus. The role of this unit in interactions with the female gametic complex is considered.