Respiratory embryology
Alain C Borczuk, MD
Dept of Pathology

Lung histology
Cast of Characters
Airways
Conducting
Respiratory
Vessels
Arteries, arterioles - pulmonary and bronchial
Capillaries
Veins/Venules and Lymphatics
Pleura- visceral and parietal

Slide 3
Lung Histology: Conducting zone
Airways Conducting Zone
Trachea
Bronchi - ciliated and goblet cells, elastic tissue, smooth muscle, glands, cartilage
Bronchioles - (1 mm) - No cartilage or bronchial glands, ciliated lining,no goblet cells, smooth muscle
Cell types
CILIATED CELL - beating of cilia contribute to mucociliary elevator
GOBLET CELL - Mucus secretion
BASAL CELL - reserve cell
KULCHITSKY CELL - neuroendocrine cells.

Pulmonary Histology
Airways Respiratory Zone
Terminal bronchiole to Respiratory bronchiole - lined by ciliated cells  and CLARA CELLS; by transitional zone to RB, all Clara cells.
Alveolar ducts/sacs
Type I cells 90% of alveolar surface
Type II cells
Cell types
CLARA CELLS - produce a component of surfactant and are the bronchiolar reserve cell
TYPE I CELLS - Thin lining cell for gas exchange
TYPE II CELLS - surfactant and alveolar reserve cell

Slide 6
Slide 7
Laryngeal development
Week 4
Respiratory primordium arises from distal/caudal pharynx
Laryngo-tracheal groove
Endodermal derivative of epithelium of larynx trachea and bronchi
Connective tissue, smooth muscle and cartilage from splanchnic mesenchyme surrounding the foregut

Laryngeal development
LT groove evaginates and forms LT diverticulum
This becomes invested with splanchnic mesoderm to form lung bud
This maintains a laryngeal inlet
The septum that forms by folds and fusion keeps a septate inlet that becomes trachea and esophagus

Epithelium of the larynx
Endoderm of proximal/cranial end of LT tube and cartilage from neural crest origin
Formation of proximal larynx – cranial tube
Arytenoid swellings grow towards tongue
Airway gets closed off, eventually recanalizes
Laryngeal webs – Incomplete recanalization
Laryngeal atresia – ascites, hydrops and lungs do not properly form.

Trachea
Endoderm of distal LT tube
Epithelium of trachea and lung
Splanchnic mesenchyme
Connective tissue
4th week
If esopahgeal separation from LT tube is incomplete, develops into TE fistula

Slide 12
Slide 13
Slide 14
Bronchi/lungs
By 28 days – endodermal buds grow along with splanchnic mesenchyme
By 35 days – Second degree bronchi, upper middle and lower on right, upper and lower on left
By 42 days – Tertiary bronchopulmonary segments, 10 on the right and 8-9 on the left.

Slide 16
Slide 17
Branching morphogenesis
By 24 weeks, 17 orders of bronchi and respiratory bronchioles (7 more after birth)
Lungs grow to pleura – visceral pleura from splanchnic mesenchyme and parietal pleura from somatic mesoderm.

Slide 19
Slide 20
Slide 21
Lung Maturation
Pseudoglandular (5-17 weeks)
No gas exchange zones
Lung resembles an exocrine gland
Canalicular (17-25 weeks)
Terminal bronchioles enlarge and branch 2-3 respiratory bronchioles then 3-6 alveolar ducts. Terminal sacs begin to form
VAscularized – caudal slower than cranial
Terminal sac (25 weeks to 34 weeks) – blood flow and surfactant
Epithelium thins to become type I like
Capillaries grow in
Blood air barrier forms
Type I and type II cells
Surfactant reduces surface tension allowing expansion.
Alveolar period (late fetal to childhood)
Surfactant
Gas exchange
Pulmonary vs systemic circulation
Alveoli mature from age 3-8. Numbers increase from 50 million at birth and 300 million at age 8 (adult number)

 Early pseudoglandular 8 wk
 Mid Pseudoglandular 13 wk
Late pseudoglandular-16 weeks
Mid-canalicular - 22 weeks
Slide 27
Slide 28
Slide 29
Congenital malformations
Cystic adenomatoid malformations
Maturation arrest in lung segments
Azyous lobe
Superior apical bronchus grows medially instead of laterally; vein is at bottom of superior lobe fissure
Sequestration –
Accessory piece of lung that becomes disconnected from tracheobronchial tree and parasitizes systemic circulation from diaphragm.

Slide 31
Breathing exercise
Begins pre-natally, allows branching to continue
Fluid is expelled from lungs at birth by vaginal pressure into capillaries and lymphatics
Fluid is needed for proper lung development
Insufficient fluid – decreased lung development
Insufficient breathing movements – decreased lung development (neurological)

Causes of Lung hypoplasia – diminished lung development
Oligohydramnios – insufficient amniotic fluid
Compression
Congenital diaphragmatic hernia – intestinal contents compress left hemithorax (usually)
Intrathoracic fluid or thoracic wall abnormality

Expansion of the lung activates a transcriptional program.
Stretching of myofibroblasts induces a transcriptional program that contributes to completion of distal proliferation and differentiation (TGF-B decrease)
Lung expansion in utero by fluid is critical to proper lung development.

RDS-Respiratory distress syndrome
Low surfactant – Respiratory distress syndrome – usually due to pre-maturity, rarely due to surfactant protein deficiency (genetic cause)
Surfactant is critical to reduce surface tension and allow lung expansion at the air fluid interface.
Inadequate surfactant leads to alveolar collapse on expiration of air, and difficulty re-inflating
Damage to the alveolus leads to cellular injury and exudation of proteins known as hyaline membranes (Hyaline membrane disease)
Continued injury from ventilation of immature lungs can lead to chronic injury known as bronchopulmonary dysplasia.
Steroids accelerate lung development and surfactant production
Surfactant can also be administered

Pulmonary vasculature
At birth, fetal lung circulation is a high pressure  that must convert to a low pressure circulation.
As air enters the lung with the first breath, oxygen tension rises.
Increased nitric oxide production increases arterial vasodilation, reducing pulmonary arterial pressure.

Molecular determinants of branching morphogenesis
Much of this data is derived from transgenic animals. Knockout of genes and gain of function mutants
Also experiments displacing mesenchyme and epithelium to new sites have been critical in understanding the crosstalk between epithelium and mesenchyma

Branching determinants
Removal of mesenchyme halts branching
Non lung mesenchyme does not support branching
Lung mesenchyme placed in trachea or salivary gland induces specific branching
Proximal vs distal mesenchyme induces site appropriate epithelial cell development
Mesenchymal factors are diffusable across membranes – No contact needed, but gradients are very local.
Epithelial factors �crosstalk� to determine mesenchymal growth and differentiation.

FGF-fibroblast growth factor
Loss of epithelial FGF receptor leads to loss of branching
FGF10 in mice from mesenchyme binds to FGFR2 on epithelial cells.
Epithelial Shh (sonic hedgehog) shuts off FGF10 from mesenchyme, stopping growth. New buds form in Shh negative areas that have persistent FGF10 production
Other FGF (e.g. FGF7) and other FGFR (e.g. FGFR3 and 4) may play a similar role in later stages of distal/alveolar lung development.

Factors in branching morphogenesis
Proliferation
Growth factors, factors that promote pluripotency
Differentiation
Transcription factors and downstream effectors leading to different cell types
Proximal and distal differences (e.g cartilage, type II cells)
Inhibition of proliferation
Growth eventually stops, also a key to branch points
Apoptosis
Structures must be remodeled in second wave of growth.
May deal with local proliferation excess.
Coordinated growth of different cell populations – �crosstalk�
Elements may be intracellular pathways, cellular receptor specificity, as well as diffusable factors with very local gradients
Differential adhesion.
Cell to cell, cell to matrix.
Adhesion molecules may be expressed reducing cellular fluidity, fostering attachment of epithelium to mesenchyme via common extracellular matrix proteins

Branching parameters
BMP4 may prevent proximal type differentiation, allowing cells to accept signals for distal development.
TTF1 and HNF3B may promote differentiation towards surfactant producing cells.
Shh may induce proliferation and differentiation in mesenchyme and inhibit epithelial proliferation signals (FGF10).
TGF-B may induce extracellular matrix production, which in turn may further anchor and stabilize epithelial cells.

Slide 42
Slide 43
Slide 44
Slide 45