Investigation of Nicotiana tabacum (+) N. suaveolens cybrids with carpelloid stamens.

To investigate cytoplasmic effects on homeotic floral morphology, Nicotiana tabacum and N. suaveolens protoplasts were fused and cybrids obtained to contrast with the sexual alloplasmic line Nta(sua)S. Nta(sua)S contains the nucleus of N. tabacum and cytoplasm of N. suaveolens while cybrids derive from fused cells where the cytoplasms can interact. The three male-sterile somatic cybrid plants analyzed contained mitochondria with N. tabacum and N. suaveolens mtDNA sequences, but not all the N. tabacum or all the N. suaveolens mtDNA sequences were present. The flowers were N. tabacum-like but with a split corolla (not observed in Nta(sua)S) and the whorl of stamens replaced by a whorl of carpel-like structures. Based on scanning electron microscopy the carpelloid stamens had a characteristic N. tabacum stigma, a style of variable length and a pseudo-ovary with ovule-like structures. The Southern blot data were consistent with mtDNA recombination. These genomic changes were maternally inherited. Chloroplasts were either of the N. tabacum or N. suaveolens type. AFLP analysis showed transfer of variable amounts of N. suaveolens nuclear DNA. However, it is the presence of the N. suaveolens sequences and/or absence of N. tabacum sequences in the mitochondria that correlates with the homeotic floral morphology. These cybrids will facilitate the analysis of the role of mitochondrial DNA sequences in floral organ identity; which has received limited attention in genetic flowering models based primarily on Arabidopsis research.

Progesterone withdrawal and estrogen activation in human parturition are coordinated by progesterone receptor A expression in the myometrium.

In human parturition, progesterone withdrawal and estrogen activation are not mediated by changes in progesterone and estrogen levels. Instead, these events could be facilitated by changes in the responsiveness of the myometrium to progesterone and estrogens via changes in PR and ER expression. We hypothesized that functional progesterone withdrawal occurs by increased expression of the type A PR (PR-A), which suppresses progesterone responsiveness, and that functional estrogen activation occurs by increased myometrial expression of ERalpha and/or ERbeta. To test this hypothesis we compared the abundance of mRNAs (assessed by quantitative RT-PCR) encoding PR-A, PR-B, ERalpha, and ERbeta in nonlaboring (n = 12) and laboring (n = 12) term human myometrium. PR-A, PR-B, the PR-A/PR-B mRNA ratio, and ERalpha mRNA were significantly increased in laboring myometrium, whereas ERbeta mRNA was low and unchanged. The PR-A/PR-B mRNA ratio correlated positively with ERalpha mRNA levels in nonlaboring myometrium and with HOXA10 mRNA levels in laboring myometrium. Because progesterone inhibits ERalpha and HOXA10 expression, these findings indicate that myometrial progesterone responsiveness is inversely related to the extent of expression of PR-A relative to PR-B. ERalpha mRNA levels correlated positively with cyclooxygenase type 2 and oxytocin receptor mRNA levels in nonlaboring myometrium, indicating that the increase in ERalpha expression is directly associated with the activation of contraction-associated genes and estrogen responsiveness. These data indicate that in the term human myometrium, responsiveness to progesterone is controlled by the expression of PR-A relative to PR-B and that a significant increase in this ratio underlies functional progesterone withdrawal. Our data also indicate that functional estrogen activation occurs by increased expression of ERalpha and is linked to functional progesterone withdrawal. Interaction between the PR and ER systems in the human myometrium may be critical for the control of human parturition and the coordination of progesterone withdrawal and estrogen activation required for parturition.