A specialized population of Periostin-expressing cardiac fibroblasts contributes to postnatal cardiomyocyte maturation and innervation

During the postnatal period in mammals, the cardiac muscle transitions from hyperplasic to hypertrophic growth, the extracellular matrix (ECM) undergoes remodeling, and the heart loses regenerative capacity. While ECM maturation and crosstalk between cardiac fibroblasts (CFs) and cardiomyocytes (CMs) have been implicated in neonatal heart development, not much is known about specialized fibroblast heterogeneity and function in the early postnatal period. In order to better understand CF functions in heart maturation and postnatal cardiomyocyte cell-cycle arrest, we have performed gene expression profiling and ablation of postnatal CF populations. Fibroblast lineages expressing Tcf21 or Periostin were traced in transgenic GFP reporter mice, and their biological functions and transitions during the postnatal period were examined in sorted cells using RNA sequencing. Highly proliferative Periostin (Postn)+ lineage CFs were found from postnatal day 1 (P1) to P11 but were not detected at P30, due to a repression ofPostngene expression. This population was less abundant and transcriptionally different from Tcf21+ resident CFs. The specialized Postn+ population preferentially expresses genes related to cell proliferation and neuronal development, while Tcf21+ CFs differentially express genes related to ECM maturation at P7 and immune crosstalk at P30. Ablation of the Postn+ CFs from P0 to P6 led to altered cardiac sympathetic nerve patterning and a reduction in binucleation and hypertrophic growth with increased fetal troponin (TroponinI1) expression in CM. Thus, postnatal CFs are heterogeneous and include a transient proliferative Postn+ population required for cardiac nerve development and cardiomyocyte maturation soon after birth.

Isolation of potential vertebrate limb-identity genes

Forelimbs and hindlimbs of tetrapods have different morphological patterns. One plausible explanation for the difference is that the cells that give rise to the limbs differentially express genes which control their pattern of development. Amphibian limb regeneration is an excellent system to test this hypothesis, since the same ultimate morphology is attained in regeneration as through embryogenesis. Using a combination of homeobox probes and differential screening, I have isolated two newt genes which are differentially expressed in regenerating forelimbs and hindlimbs. One of these genes displays properties expected of a gene involved in controlling limb morphology, including expression in mesodermal tissue and constancy of expression upon transplantation. Based on sequence analysis, this gene appears to be homologous to a homeobox-containing gene previously isolated from frog and human libraries.